disc cell phenotypes: Topics by
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Our objectives were to use transmission electron microscopy to analyze more fully inclusion bodies in the annulus pulposus and to study the extracellular matrix ECM surrounding cells containing inclusion bodies. Application Area: Analyze cell structure and morphology.
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Inclusion bodies have been identified previously in chondrocytes and osteoblasts in chondrodysplasias and osteogenesis imperfecta. Inclusion bodies in intervertebral disc cells , however, have only recently been recognized. Our objectives were to use transmission electron microscopy to analyze more fully inclusion bodies in the annulus pulposus and to study the extracellular matrix ECM surrounding cells containing inclusion bodies.
ECM frequently encapsulated cells with inclusion bodies, and commonly contained prominent banded aggregates of Type VI collagen. Inclusion body material had several morphologies, including relatively smooth, homogeneous material, or a rougher, less homogeneous feature.
Such findings expand our knowledge of the fine structure of the human disc cell and ECM during disc degeneration, and indicate the potential utility of ultrastructural identification of discs with intracellular inclusion bodies as a screening method for molecular studies directed toward identification of defective gene products in degenerating discs.
Human umbilical cord mesenchymal stromal cells exhibit immature nucleus pulposus cell phenotype in a laminin-rich pseudo-three-dimensional culture system. Cell supplementation to the herniated or degenerated intervertebral disc IVD is a potential strategy to promote tissue regeneration and slow disc pathology. Human umbilical cord mesenchymal stromal cells HUCMSCs – originating from the Wharton’s jelly – remain an attractive candidate for such endeavors with their ability to differentiate into multiple lineages.
Previously, mesenchymal stem cells MSCs have been studied as a potential source for disc tissue regeneration. However, no studies have demonstrated that MSCs can regenerate matrix with unique characteristics matching that of immature nucleus pulposus NP tissues of the IVD. In our prior work, immature NP cells were found to express specific laminin isoforms and laminin-binding receptors that may serve as phenotypic markers for evaluating MSC differentiation to NP-like cells.
The goal of this study is to evaluate these markers and matrix synthesis for HUCMSCs cultured in a laminin-rich pseudo-three-dimensional culture system. Cells were cultured under hypoxia environment with three differentiation conditions: NP differentiation media containing 2.
Cell clustering behavior, matrix production and the expression of NP-specific laminin and laminin-receptors were evaluated at days 1, 7, 13 and 21 of culture. Data show that a pseudo-three-dimensional culture condition laminin-1 rich promoted HUCMSC differentiation under no serum conditions. Starting at day 1, HUCMSCs demonstrated a cell clustering morphology similar to that of immature NP cells in situ and that observed for primary immature NP cells within the similar laminin-rich culture system prior study.
Human disc cells in monolayer vs 3D culture: cell shape, division and matrix formation. Full Text Available Abstract Background The relationship between cell shape, proliferation, and extracellular matrix ECM production, important aspects of cell behavior, is examined in a little-studied cell type, the human annulus cell from the intervertebral disc , during monolayer vs three-dimensional 3D culture. Results Three experimental studies showed that cells respond specifically to culture microenvironments by changes in cell shape, mitosis and ECM production: 1 Cell passages showed extensive immunohistochemical evidence of Type I and II collagens only in 3D culture.
Chondroitin sulfate and keratan sulfate were abundant in both monolayer and 3D cultures. Conclusions The cell ‘s in vivo interactions with the ECM can regulate shape, gene expression and other cell functions. The shape of the annulus cell changes markedly during life: the young, healthy disc contains spindle shaped cells and abundant collagen.
With aging and degeneration, many cells assume a strikingly different appearance, become rounded and are surrounded by unusual accumulations of ECM products. In vitro manipulation of disc cells provides an experimental window for testing how disc cells from given individuals respond when they are grown in environments which direct cells to have either spindle- or rounded-shapes.
These findings contribute new information to the important issue of the influence of cell shape on cell behavior. Degenerative disc disease DDD of the cervical spine is common after middle age and can cause loss of disc height with painful nerve impingement, bone and joint inflammation.
Despite the clinical importance of these problems, in current publications the pathology of cervical disc degeneration has been studied merely from a morphologic view point using magnetic resonance imaging MRI , without addressing the issue of biological treatment approaches.
So far a wide range of endogenously expressed bioactive factors in degenerative cervical disc cells has not yet been investigated, despite its importance for gene therapeutic approaches. Although degenerative lumbar disc cells have been targeted by different biological treatment approaches, the quantities of disc cells and the concentrations of gene therapeutic factors used in animal models differ extremely. These indicate lack of experimentally acquired data regarding disc cell proliferation and levels of target proteins.
Therefore, we analysed proliferation and endogenous expression levels of anabolic, catabolic, ant-catabolic, inflammatory cytokines and matrix proteins of degenerative cervical disc cells in three-dimensional cultures. NP cells were cultured for four weeks with low-glucose in collagen I scaffold. Their proliferation rates were analysed using 3- 4, 5-dimethylthiazolyl-2 -2,5-diphenyltetrazolium bromide. Their protein expression levels of 28 therapeutic targets were analysed using enzyme-linked immunosorbent assay.
During progressive grades of degeneration NP cell proliferation rates were similar. Significantly decreased aggrecan and collagen II expressions P Disc size regulation in the brood cell building behavior of leaf-cutter bee, Megachile tsurugensis.
The leaf-cutter bee, Megachile tsurugensis, builds a brood cell in a preexisting tunnel with leaf discs that she cuts in decreasing sizes and assembles them like a Russian matryoshka doll.
By experimentally manipulating the brood cell , it was investigated how she regulates the size of leaf discs that fit in the brood cell ‘s internal volume. When the internal volume was artificially increased by removing a bulk of leaf discs , she decreased the leaf disc size, although increasing it would have made the leaf disc more fitting in the increased internal volume. As a reverse manipulation, when the internal volume was decreased by inserting a group of inner layers of preassembled leaf discs to a brood cell , she decreased the leaf disc size, so that the leaf disc could fit in the decreased internal volume.
These results suggest that she uses at least two different mechanisms to regulate the disc size: the use of some internal memory about the degree of building work accomplished in the first and of sensory feedback of dimensional information at the construction site in the second manipulation, respectively. It was concluded that a stigmergic mechanism, an immediate sensory feedback from the brood cell changed by the building work, alone cannot explain the details of the bee’s behavior particularly with respect to her initial response to the first manipulation.
For a more complete explanation of the behavior exhibited by the solitary bee, two additional behavioral elements, reinforcement of building activity and processing of dimensional information, were discussed along with stigmergy. Full Text Available Much emphasis has been placed recently on the repair of degenerate discs using implanted cells , such as disc cells or bone marrow derived mesenchymal stem cells MSCs.
This study examines the temporal response of bovine and human nucleus pulposus NP cells and MSCs cultured in monolayer following exposure to altered levels of glucose 0, 3. NP cells were also exposed to the cell death inducers, hydrogen peroxide and staurosporine, in comparison to serum starvation. However, as time progresses, NP cells supplemented with serum recover with minimal evidence of cell death.
Human NP cells show no evidence of proliferation in response to nutrient supplementation, whereas MSCs showed greater response to increased nutrition. When specifically inducing NP cell death with hydrogen peroxide and staurosporine, as expected, the cell number declined.
These results support the concept that implanted NP cells or MSCs may be capable of survival in the nutrient-poor environment of the degenerate human disc , which has important clinical implications for the development of IVD cell therapies. The design and fabrication of a heating system has been a significant challenge in implementing chemical lysis on a lab-on-a- disc LOD.
The proposed system contains a sample inlet, phase change material PCM array, heating chamber, and valve in a single disc , providing cost-effective, rapid, and fully automated chemical cell lysis.
Compared to the conventional cell lysis system, our cell lysis system has many advantages, such as a compact structure that is easily integrated into the LOD and reduced processing time and labor. The experiments are conducted with Salmonella typhimurium strains to demonstrate the performance.
The experimental results show that the proposed approach is greatly effective in realizing a chemical cell lysis system on an LOD with higher throughput in terms of purity and yield of DNA. The inhibition zone diameters were measured and interpreted at 7 h for the presence of carbapenemase.
This early phenotypic method is very simple, inexpensive, and reliable in the detection and differentiation of carbapenemase-producing Enterobacteriaceae. It could be exploited in any microbiological laboratory for diagnosis of these recalcitrant bacteria. This assay poses excellent performance in discrimination of Kl. The rapid detection could help clinicians screen patients, control infection and provide epidemiological surveillance. Results therefore suggest that successful treatments must promote anabolism and cell proliferation in.
Stem- cell treatment in disc degeneration: What is the evidence? Full Text Available To review the potential role of stem cells in treating degenerative disc disease of the intervertebral disc IVD. A review was performed of articles from the Medline database concerning stem cells and degenerative disc disease DDD.
To discuss the data, the papers were classified as: review, in vitro, experimental, and clinical. The currently available treatments were basically for symptom reduction, not to revert the IVD degenerative process.
Besides, experimental studies demonstrated that the MSC remained viable when injected into the IVD, and that they were able to regenerate partially from the degenerated IVD and its structure. The few clinical studies found in the literature presented diverging results. Although many advances are being achieved in studies in vitro and experimental, there is a lack of clinical studies to prove the role of MSC in DDD management.
Lactoferricin enhances BMP7-stimulated anabolic pathways in intervertebral disc cells. Bone-morphogenetic protein-7 BMP7 is a well-known anabolic and anti-catabolic growth factor on intervertebral disc IVD matrix and cell homeostasis.
In this study, we investigated the potential benefits of using combined peptide therapy with LfcinB and BMP7 for intervertebral disc matrix repair and to understand cellular and signaling mechanisms controlled by these factors. We studied the effects of BMP7 and LfcinB as individual treatments and combined therapy on bovine nucleus pulposus NP cells by assessing proteoglycan PG accumulation and synthesis, and the gene expression of matrix protein aggrecan and transcription factor SOX Treatment of bovine NP cells cultured in alginate with LfcinB plus BMP7 synergistically stimulates PG synthesis and accumulation in part by upregulation of aggrecan gene expression.
These data indicate that LfcinB-suppression of Noggin may eliminate the negative feedback of BMP7, thereby maximizing biological activity of BMP7 and ultimately shifting homeostasis to a pro-anabolic state in disc cells.
We propose that combination growth factor therapy using BMP7 and LfcinB may be beneficial for treatment of disc degeneration.
The effects of oxygen level and glucose concentration on the metabolism of porcine TMJ disc cells. To determine the combined effect of oxygen level and glucose concentration on cell viability, ATP production, and matrix synthesis of temporomandibular joint TMJ disc cells. TMJ disc cells were isolated from pigs aged months and cultured in a monolayer.
Cell cultures were preconditioned for 48 h with 0, 1. The cell viability was measured using the WST-1 assay. ATP production was determined using the Luciferin-Luciferase assay. Collagen and proteoglycan synthesis were determined by measuring the incorporation of [2, 3- 3 H] proline and [ 35 S] sulfate into the cells , respectively. TMJ disc cell viability significantly decreased P oxygen levels significantly increased viability P oxygen levels significantly reduced ATP production P oxygen was significant in regards to cell viability P oxygen are important, glucose is the limiting nutrient for TMJ disc cell survival.
At low oxygen levels, the production of ATP, collagen, and proteoglycan are severely inhibited. These results suggest that steeper nutrient gradients may exist in the TMJ disc and it may be vulnerable to pathological events that impede nutrient supply. Published by Elsevier Ltd. Using a preclinical genetic approach, we investigated whether simultaneous disruption of the risk factors Neuregulin-1 NRG1 and Disrupted-in-schizophrenia 1 DISC 1 would produce a disease-relevant phenotypic profile different from that observed following disruption to either gene alone.
NRG1 heterozygotes exhibited hyperactivity and disruption to prepulse inhibition, both reversed by antipsychotic treatment, and accompanied by reduced striatal dopamine D2 receptor protein expression, impaired social cognition, and altered glutamatergic synaptic protein expression in selected brain areas.
Single gene DISC 1 mutants demonstrated a disruption in social cognition and nest-building, altered brain 5-hydroxytryptamine levels and hippocampal ErbB4 expression, and decreased cortical expression of the schizophrenia-associated microRNA miRb.
For permissions, please email: journals. Patterning and growth are linked during early development and have to be tightly controlled to result in a functional tissue or organ. During the development of the Drosophila eye, this linkage is particularly clear: the growth of the eye primordium mainly results from proliferating cells ahead of the morphogenetic furrow MF , a moving signaling wave that sweeps across the tissue from the posterior to the anterior side, that induces proliferating cells anterior to it to differentiate and become cell cycle quiescent in its wake.
Therefore, final eye disc size depends on the proliferation rate of undifferentiated cells and on the speed with which the MF sweeps across the eye disc. We developed a spatio-temporal model of the growing eye disc based on the regulatory interactions controlled by the signals Decapentaplegic Dpp , Hedgehog Hh and the transcription factor Homothorax Hth and explored how the signaling patterns affect the movement of the MF and impact on eye disc growth.
We used published and new quantitative data to parameterize the model. In particular, two crucial parameter values, the degradation rate of Hth and the diffusion coefficient of Hh, were measured.
The model is able to reproduce the linear movement of the MF and the termination of growth of the primordium. We further show that the model can explain several mutant phenotypes , but fails to reproduce the previously observed scaling of the Dpp gradient in the anterior compartment. Full Text Available Patterning and growth are linked during early development and have to be tightly controlled to result in a functional tissue or organ.
During the development of the Drosophila eye, this linkage is particularly clear: the growth of the eye primordium mainly results from proliferating cells ahead of the morphogenetic furrow MF, a moving signaling wave that sweeps across the tissue from the posterior to the anterior side, that induces proliferating cells anterior to it to differentiate and become cell cycle quiescent in its wake.
We developed a spatio-temporal model of the growing eye disc based on the regulatory interactions controlled by the signals Decapentaplegic Dpp, Hedgehog Hh and the transcription factor Homothorax Hth and explored how the signaling patterns affect the movement of the MF and impact on eye disc growth. Cell therapy for intervertebral disc repair: advancing cell therapy from bench to clinics.
Full Text Available Intervertebral disc IVD degeneration is a major cause of pain and disability; yet therapeutic options are limited and treatment often remains unsatisfactory. In recent years, research activities have intensified in tissue engineering and regenerative medicine, and pre-clinical studies have demonstrated encouraging results.
Nonetheless, the translation of new biological therapies into clinical practice faces substantial barriers. During the symposium “Where Science meets Clinics”, sponsored by the AO Foundation and held in Davos, Switzerland, from September , , hurdles for translation were outlined, and ways to overcome them were discussed. With respect to cell therapy for IVD repair, it is obvious that regenerative treatment is indicated at early stages of disc degeneration, before structural changes have occurred.
It is envisaged that in the near future, screening techniques and non-invasive imaging methods will be available to detect early degenerative changes. The promises of cell therapy include a sustained effect on matrix synthesis, inflammation control, and prevention of angio- and neuro-genesis.
Discogenic pain, originating from “black discs ” or annular injury, prevention of adjacent segment disease, and prevention of post-discectomy syndrome were identified as prospective indications for cell therapy. Before such therapy can safely and effectively be introduced into clinics, the identification of the patient population and proper standardisation of diagnostic parameters and outcome measurements are indispensable.
Furthermore, open questions regarding the optimal cell type and delivery method need to be resolved in order to overcome the safety concerns implied with certain procedures. Finally, appropriate large animal models and well-designed clinical studies will be required, particularly addressing safety aspects.
Primary immune system responders to nucleus pulposus cells : evidence for immune response in disc herniation. Full Text Available Although intervertebral disc herniation and associated sciatica is a common disease, its molecular pathogenesis is not well understood.
Immune responses are thought to be involved. This study provides direct evidence that even non-degenerated nucleus pulposus NP cells elicit immune responses. An in vitro colony forming inhibition assay demonstrated the suppressive effects of autologous spleen cells on NP cells and an in vitro cytotoxicity assay showed the positive cytotoxic effects of natural killer NK cells and macrophages on NP cells.
Non-degenerated rat NP tissues transplanted into wild type rats and immune-deficient mice demonstrated a significantly higher NP cell survival rate in immune-deficient mice. Immunohistochemical staining showed the presence of macrophages and NK cells in the transplanted NP tissues.
These results suggest that even non-degenerated autologous NP cells are recognized by macrophages and NK cells , which may have an immunological function in the early phase of disc herniation. These findings contribute to understanding resorption and the inflammatory reaction to disc herniation.
The generation and functional characterization of induced pluripotent stem cells from human intervertebral disc nucleus pulposus cells. Disc degenerative disease DDD is believed to originate in the nucleus pulposus NP region therefore, it is important to obtain a greater number of active NP cells for the study and therapy of DDD.
Human induced pluripotent stem cells iPSCs are a powerful tool for modeling the development of DDD in humans, and have the potential to be applied in regenerative medicine. Successful reprogramming of iPSCs was verified by the expression of surface markers and presence of teratoma. Differentiation of iPSCs into NP-like cells was performed in a culture plate or in hydrogel, whereby skin fibroblast derived-iPSCs were used as a control.
Results demonstrated that iPSCs derived from NP cells displayed a normal karyotype, expressed pluripotency markers, and formed teratoma in nude mice. This method provides a novel treatment of DDD by using patient-specific NP cells in a relatively simple and straightforward manner.
Degenerative disc disease DDD is associated with spinal pain often leading to long-term disability. However, the non-chondrodystrophic canine intervertebral disc is protected from the development of DDD, ostensibly due to its retention of notochordal cells NC in the nucleus pulposus NP.
In this study, we hypothesized that secretome analysis of the NC-rich NP will lead to the identification of key proteins that delay the onset of DDD. Increased osmolarity and cell clustering preserve canine notochordal cell phenotype in culture. Spillekom, S. Degeneration of the intervertebral disc IVD is associated with a loss of notochordal cells NCs from the nucleus pulposus NP and their replacement by chondrocyte-like cells.
NCs are known to maintain extracellular matrix quality and stimulate the chondrocyte-like NP cells , making NCs attractive. DISC 1 and striatal volume: a potential risk phenotype for mental illness. Full Text Available Disrupted-in-schizophrenia 1 was originally discovered in a large Scottish family with abnormally high rates of severe mental illness, including schizophrenia, bipolar disorder, and depression.
An accumulating body of evidence from genetic, postmortem, and animal data supports a role for DISC 1 in different forms of mental illness. DISC 1 may play an important role in determining structure and function of several brain regions. One brain region of particular importance for several mental disorders is the striatum, and DISC 1 mutant mice have demonstrated an increase in dopamine D2 receptors in this structure. However, association between DISC 1 functional polymorphisms and striatal structure have not been examined in humans to our knowledge.
We, therefore hypothesized that there would be a relationship between human striatal volume and DISC 1 genotype, specifically in the LeuPhe rs and SerCys rs single nucleotide polymorphisms.
We tested our hypothesis by automatically identifying the striatum in fifty-four healthy volunteers recruited for this study. We also performed an exploratory analysis of cortical thickness, cortical surface area, and structure volume.
However, these exploratory findings do not survive a conservative correction for multiple comparisons. Our findings demonstrate that a functional DISC 1 variant influences striatal volumes. Full Text Available Low back pain and degenerative disc disease are a significant cause of pain and disability worldwide.
Advances in regenerative medicine and cell -based therapies, particularly the transplantation of mesenchymal stem cells and intervertebral disc chondrocytes, have led to the publication of numerous studies and clinical trials utilising these biological therapies to treat degenerative spinal conditions, often reporting favourable outcomes.
Stem cell mediated disc regeneration may bridge the gap between the two current alternatives for patients with low back pain, often inadequate pain management at one end and invasive surgery at the other. Through cartilage formation and disc regeneration or via modification of pain pathways stem cells are well suited to enhance spinal surgery practice. This paper will systematically review the current status of basic science studies, preclinical and clinical trials utilising cell -based therapies to repair the degenerate intervertebral disc.
The mechanism of action of transplanted cells , as well as the limitations of published studies, will be discussed. Adipose stem cells for intervertebral disc regeneration: Current status and concepts for the future: Tissue Engineering Review Series. Introduction Degenerative disc disease and emerging biological treatment approaches Stem cell sources Integration of ASC-based regenerative medicine and surgery In vitro studies Animal models Cells in disc regeneration in vivo In vivo studies Perspective Conclusions Abstract New regenerative.
A potential role for cell -based therapeutics in the treatment of intervertebral disc herniation. Lower back pain and disc degeneration negatively affect quality of life and impose an enormous financial burden.
An extensive body of scientific work has evolved that characterizes the disc , demonstrating spinal anatomy and morphology that contribute to risk and likely promote failure. Ultimately, matrix failure is responsible for mechanical failure, which in turn results in spinal compromise anatomically and subsequent pain.
One intervening approach to breaking this sequence has been to repopulate the anatomy with autologous disc chondrocytes– cells capable of restoring the matrix and retaining the mechanical balance by which the disc functions.
This strategy has been implemented both in patients and in animal models, and early results, although preliminary, support the premise as a positive approach. A mathematical model of cancer cells with phenotypic plasticity.
Full Text Available Purpose: The phenotypic plasticity of cancer cells is recently becoming a cutting-edge research area in cancer, which challenges the cellular hierarchy proposed by the conventional cancer stem cell theory.
In this study, we establish a mathematical model for describing the phenotypic plasticity of cancer cells , based on which we try to find some salient features that can characterize the dynamic behavior of the phenotypic plasticity especially in comparison to the hierarchical model of cancer cells. Methods: We model cancer as population dynamics composed of different phenotypes of cancer cells.
In this model, not only can cancer cells divide symmetrically and asymmetrically and die, but they can also convert into other cellular phenotypes. According to the Law of Mass Action, the cellular processes can be captured by a system of ordinary differential equations ODEs. On one hand, we can analyze the long-term stability of the model by applying qualitative method of ODEs.
On the other hand, we are also concerned about the short-term behavior of the model by studying its transient dynamics. Meanwhile, we validate our model to the cell -state dynamics in published experimental data. Results: Our results show that the phenotypic plasticity plays important roles in both stabilizing the distribution of different phenotypic mixture and maintaining the cancer stem cells proportion. Conclusion: Since the validity of the phenotypic plasticity paradigm and the conventional cancer stem cell theory is still debated in experimental biology, it is worthy of theoretically searching for good indicators to distinguish the two models through quantitative methods.
According to our study, the phenotypic equilibrium and overshoot. Bovine annulus fibrosus cell lines isolated from intervertebral discs. Full Text Available The adult bovine Bos taurus intervertebral disc is primarily comprised of two major tissue types: The outer annulus fibrosus AF and the central nucleus pulposus NP.
We isolated several primary cell lineages of passage P 0 cells from the AF tissue omitting typically used enzymatic tissue digestion protocols. Preliminary analysis of the AF derived cells for expression of the two structural genes Col1a1 and Col2a1 was performed by PISH recapitulating the expression observed in vivo. A computational study of intervertebral disc degeneration in relation to changes in regional tissue composition and disc nutrition.
Altered disc cell nutrition affects cell viability and can generate catabolic cascades that degrade the extracellular matrix ECM. Also, a major degenerative biochemical change in the disc is the proteoglycan PG loss, which affects the osmotic pressure and hydration that is critical for cell nutrition. However, the relationship between biochem Sonic hedgehog in the notochord is sufficient for patterning of the intervertebral discs. The intervertebral discs , located between adjacent vertebrae, are required for stability of the spine and distributing mechanical load throughout the vertebral column.
All cell types located in the middle regions of the discs , called nuclei pulposi, are derived from the embryonic notochord. Recently, it was shown that the hedgehog signaling pathway plays an essential role during formation of nuclei pulposi. However, during the time that nuclei pulposi are forming, Shh is expressed in both the notochord and the nearby floor plate.
To determine the source of SHH protein sufficient for formation of nuclei pulposi we removed Shh from either the floor plate or the notochord using tamoxifen-inducible Cre alleles.
Removal of Shh from the floor plate resulted in phenotypically normal intervertebral discs , indicating that Shh expression in this tissue is not required for disc patterning. In addition, embryos that lacked Shh in the floor plate had normal vertebral columns, demonstrating that Shh expression in the notochord is sufficient for pattering the entire vertebral column. Removal of Shh from the notochord resulted in the absence of Shh in the floor plate, loss of intervertebral discs and vertebral structures.
These data indicate that Shh expression in the notochord is sufficient for patterning of the intervertebral discs and the vertebral column. Increased density of DISC 1-immunoreactive oligodendroglial cells in fronto-parietal white matter of patients with paranoid schizophrenia. Profound white matter abnormalities have repeatedly been described in schizophrenia, which involve the altered expression of numerous oligodendrocyte-associated genes.
Transcripts of the disrupted-in-schizophrenia 1 DISC 1 gene, a key susceptibility factor in schizophrenia, have recently been shown to be expressed by oligodendroglial cells and to negatively regulate oligodendrocyte differentiation and maturation. To learn more about the putative role s of oligodendroglia-associated DISC 1 in schizophrenia, we analyzed the density of DISC 1-immunoreactive oligodendrocytes in the fronto-parietal white matter in postmortem brains of patients with schizophrenia.
Pathophysiologically, over-expression of DISC 1 protein s in white matter oligodendrocytes might add to the reduced levels of two myelin markers, 2′,3′-cyclic-nucleotide 3′-phosphodiesterase and myelin basic protein in schizophrenia. Moreover, it might significantly contribute to cell cycle abnormalities as well as to deficits in oligodendroglial cell differentiation and maturation found in schizophrenia. Multiparametric classification links tumor microenvironments with tumor cell phenotype.
Full Text Available While it has been established that a number of microenvironment components can affect the likelihood of metastasis, the link between microenvironment and tumor cell phenotypes is poorly understood.
Here we have examined microenvironment control over two different tumor cell motility phenotypes required for metastasis. By high-resolution multiphoton microscopy of mammary carcinoma in mice, we detected two phenotypes of motile tumor cells , different in locomotion speed. Only slower tumor cells exhibited protrusions with molecular, morphological, and functional characteristics associated with invadopodia. Each region in the primary tumor exhibited either fast- or slow-locomotion.
To understand how the tumor microenvironment controls invadopodium formation and tumor cell locomotion, we systematically analyzed components of the microenvironment previously associated with cell invasion and migration. No single microenvironmental property was able to predict the locations of tumor cell phenotypes in the tumor if used in isolation or combined linearly.
To solve this, we utilized the support vector machine SVM algorithm to classify phenotypes in a nonlinear fashion. This approach identified conditions that promoted either motility phenotype. We then demonstrated that varying one of the conditions may change tumor cell behavior only in a context-dependent manner. In addition, to establish the link between phenotypes and cell fates, we photoconverted and monitored the fate of tumor cells in different microenvironments, finding that only tumor cells in the invadopodium-rich microenvironments degraded extracellular matrix ECM and disseminated.
The number of invadopodia positively correlated with degradation, while the inhibiting metalloproteases eliminated degradation and lung metastasis, consistent with a direct link among invadopodia, ECM degradation, and metastasis. We have detected and characterized two phenotypes of motile tumor cells in vivo, which. Bone-morphogenetic protein-7 BMP-7 is a growth factor that plays a major role in mediating anabolism and anti-catabolism of the intervertebral disc matrix and cell homeostasis.
However, the role FoxC2 may play in degenerative human intervertebral disc tissue and the relationship between FoxC2 and BMP-7 in nucleus pulposus NP cells remain to be elucidated.
Phenotypic and functional plasticity of cells of innate immunity: macrophages, mast cells and neutrophils. Hematopoietic cells , including lymphoid and myeloid cells , can develop into phenotypically distinct ‘subpopulations’ with different functions. However, evidence indicates that some of these subpopulations can manifest substantial plasticity that is, undergo changes in their phenotype and function Here we focus on the occurrence of phenotypically distinct subpopulations in three lineages of myeloid cells with important roles in innate and acquired immunity: macrophages, mast cells and neutrophils.
Cytokine signals, epigenetic modifications and other microenvironmental factors can substantially This suggests that regulation of the phenotype and function of differentiated hematopoietic cells by microenvironmental factors, including those generated during immune responses, represents Biomaterials for intervertebral disc regeneration and repair. The intervertebral disc contributes to motion, weight bearing, and flexibility of the spine, but is susceptible to damage and morphological changes that contribute to pathology with age and injury.
Engineering strategies that rely upon synthetic materials or composite implants that do not interface with the biological components of the disc have not met with widespread use or desirable outcomes in the treatment of intervertebral disc pathology. Here we review bioengineering advances to treat disc disorders, using cell -supplemented materials, or acellular, biologically based materials, that provide opportunity for cell -material interactions and remodeling in the treatment of intervertebral disc disorders.
While a field still in early development, bioengineering-based strategies employing novel biomaterials are emerging as promising alternatives for clinical treatment of intervertebral disc disorders. The effect of novel nitrogen-rich plasma polymer coatings on the phenotypic profile of notochordal cells. Full Text Available Abstract Background The loss of the notochordal cells from the nucleus pulposus is associated with ageing and disc degeneration.
However, understanding the mechanisms responsible for the loss of these cells has been hampered in part due to the difficulty of culturing and maintaining their phenotype. Furthermore, little is known about the influence of the substratum on the molecular markers of notochordal cells.
Results Notochordal cells were maintained in culture on PPE:N for up to 14 days with no loss in cell viability. Except for VIM, gene expression varied depending on the culture periods and 23971 concentration of the substratum. Generally, PPE:N surfaces altered gene expression significantly when cells were cultured for 3 or 7 days.
Conclusion The present study has shown that notochordal cells from dogs can attach to and grow on PPE:N surfaces. Analysis of the expression of different genes in these cells cultured on different N-functionalized surfaces indicates that cellular behaviour is gene-specific and time-dependent. Further studies are required to better understand the roles of specific surface functionalities on receptor sites, and their effects on cellular phenotypes.
DISC 1 disrupted-in-schizophrenia-1 regulates differentiation of oligodendrocytes. Here we report that mammalian DISC 1 endogenously expressed in oligodendroglial lineage cells negatively regulates differentiation of oligodendrocyte precursor cells into oligodendrocytes.
DISC 1 mRNA was expressed in primary cultured rat cortical oligodendrocyte precursor cells and decreased when oligodendrocyte precursor cells were induced to differentiate by PDGF deprivation. Immunocytochemical analysis showed that overexpressed DISC 1 was localized in the cell bodies and processes of oligodendrocyte precursor cells and oligodendrocytes.
We show that expression of the myelin related markers, CNPase and MBP, as well as the number of cells with a matured oligodendrocyte morphology, were decreased following full length DISC 1 overexpression. Overexpression of a truncated form of DISC 1 also resulted in an increase in expression of myelin related proteins and the number of mature oligodendrocytes, potentially acting via a dominant negative mechanism. We also identified involvement of Sox10 and Nkx2.
Rabbit model of intervertebral disc degeneration by external compression device characterized by X-ray, MRI, histology, and cell viability. Full Text Available Appropriate experimental animal models, which mimic the degenerative process occurring in human intervertebral disc IVD breakdown and can be used for new treatment studies such as tissue engineering or disc distraction are lacking.
We studied the external compression device that used by Kroeber et al to create intervertebral disc degeneration in rabbit model characterized by X-ray, MRI, Histology, and Cell Viability. Ten NZW rabbit were randomly assigned to one of five groups. Intervertebral disc VL4-L5 are compressed using an external loading device, 1.
First group rabbit are loaded for 14 days, second loaded for 28 days, thirth group are loaded for 14 days, and unloaded for 14 days, fourth group loaded for 28 days and unloaded for 28 days. The fifth group, rabbits underwent a sham operation. Additional, rabbits were used as sample for cell viability study. In disc height : sample in group one have biggest decreasing of disc height, that is In MRI assessment, the worst grade is grade 3. In histological score, the worst group is group three Group one have the largest dead cell , that are Trypan blue staining showed that group four have better viable cell The study conclude disc degeneration can be created by external axial loading for 14 days in rabbit intervertebral disc.
Duration of 28 days unloading gave better result for cells to recover. Aberrant phenotypes in peripheral T cell lymphomas. Seventy six peripheral T cell lymphomas were examined immunohistologically to test their reactivity with a panel of monoclonal antibodies against 11 T cell associated antigens CD, CD27, UCHL1, and the T cell antigen receptor.
No correlation was seen between the occurrence of aberrant phenotypes and the histological subtype. It is concluded that the demonstration of an aberrant phenotype is a valuable supplement to histological assessment in the diagnosis of peripheral T cell lymphomas.
It is recommended that the panel of monoclonal antibodies against T cell differentiation antigens should be fairly large, as apparently any antigen may be lost in the process of malignant transformation. Structural phenotyping of stem cell -derived cardiomyocytes.
Structural phenotyping based on classical image feature detection has been adopted to elucidate the molecular mechanisms behind genetically or pharmacologically induced changes in cell morphology. Here, we developed a set of 11 metrics to capture the increasing sarcomere organization that occurs intracellularly during striated muscle cell development. Further, we combined these metrics with data mining algorithms to unbiasedly score the phenotypic maturity of human-induced pluripotent stem cell -derived cardiomyocytes.
Published by Elsevier Inc. Full Text Available The direct methanol fuel cell DMFC adopts methanol solution as a fuel suitable for low power portable applications. A miniature, lightweight, passive air-breathing design is therefore desired. This paper presents a novel planar disc -type DMFC with multiple cells containing a novel developed lightweight current collector at both the anode and cathode sides. The current collecting areas are fabricated by sequentially coating a corrosion resistant layer and electrical conduction layer via the thermal evaporation technique.
The anode current collector has carved flow channels for fuel transport and production. The cathode current collector has drilled holes for passive air breathing. In order to ensure feasibility in the present concept a 3- cell prototype DMFC module with lightweight disc type current collectors is designed and constructed.
Experiments were conducted to measure the cell performance. The results show that the highest cell power output is Phenotypic equilibrium as probabilistic convergence in multi- phenotype cell population dynamics. Full Text Available We consider the cell population dynamics with n different phenotypes. Both the Markovian branching process model stochastic model and the ordinary differential equation ODE system model deterministic model are presented, and exploited to investigate the dynamics of the phenotypic proportions.
We will prove that in both models, these proportions will tend to constants regardless of initial population states ” phenotypic equilibrium” under weak conditions, which explains the experimental phenomenon in Gupta et al. We also prove that Gupta et al. As an application, we will give sufficient and necessary conditions under which the proportion of one phenotype tends to 0 die out or 1 dominate.
We also extend our results to non-Markovian cases. Human disc degeneration is associated with increased MMP 7 expression. During intervertebral disc IVD degeneration, normal matrix synthesis decreases and degradation of disc matrix increases.
A number of proteases that are increased during disc degeneration are thought to be involved in its pathogenesis. To date, however, it is not known how its expression changes with degeneration or its exact location. We investigated the localization of MMP 7 in human, histologically graded, nondegenerate, degenerated and prolapsed discs to ascertain whether MMP 7 is up-regulated during disc degeneration.
Samples of human IVD tissue were fixed in neutral buffered formalin, embedded in paraffin, and sections stained with hematoxylin and eosin to score the degree of morphological degeneration. We found that the chondrocyte-like cells of the nucleus pulposus and inner annulus fibrosus were MMP 7 immunopositive; little immunopositivity was observed in the outer annulus.
Nondegenerate discs showed few immunopositive cells. A significant increase in the proportion of MMP 7 immunopositive cells was seen in the nucleus pulposus of discs classified as showing intermediate levels of degeneration and a further increase was seen in discs with severe degeneration. Prolapsed discs showed more MMP 7 immunopositive cells compared to nondegenerated discs , but fewer than those seen in cases of severe degeneration.
Induction of appropriate Th- cell phenotypes : cellular decision-making in heterogeneous environments. Helper T Th – cell differentiation is a key event in the development of the adaptive immune response. By the production of a range of cytokines, Th cells determine the type of immune response that is raised against an invading pathogen.
Th cells can adopt many different phenotypes , and Th- cell phenotype decision-making is crucial in mounting effective host responses. This review discusses the different Th- cell phenotypes that have been identified and how Th cells adopt a particular phenotype. The regulation of Th- cell phenotypes has been studied extensively using mathematical models, which have explored the role of regulatory mechanisms such as autocrine cytokine signalling and cross-inhibition between self-activating transcription factors.
At the single cell level, Th responses tend to be heterogeneous, but corrections can be made soon after T- cell activation. Although pathogens and the innate immune system provide signals that direct the induction of Th- cell phenotypes , these instructive mechanisms could be easily subverted by pathogens.
We discuss that a model of success-driven feedback would select the most appropriate phenotype for clearing a pathogen. Given the heterogeneity in the induction phase of the Th response, such a success-driven feedback loop would allow the selection of effective Th- cell phenotypes while terminating incorrect responses.
Effects of age, replicative lifespan and growth rate of human nucleus pulposus cells on selecting age range for cell -based biological therapies for degenerative disc diseases. Autologous disc cell implantation, growth factors and gene therapy appear to be promising therapies for disc regeneration.
Unfortunately, the replicative lifespan and growth kinetics of human nucleus pulposus NP cells related to host age are unclear. We investigated the potential relations among age, replicative lifespan and growth rate of NP cells , and determined the age range that is suitable for cell -based biological therapies for degenerative disc diseases.
We used NP tissues classified by decade into five age groups: 30s, 40s, 50s, 60s and 70s. The mean cumulative PDL and PDR in the younger groups 30s, 40s and 50s were significantly higher than those in the older groups 60s and 70s.
We found that the replicative lifespan and growth rate of human NP cells decreased with age. The replicative potential of NP cells decreased significantly in patients 60 years old and older. Young individuals less than 60 years old may be suitable candidates for NP cell -based biological therapies for treating degenerative disc diseases.
Total disc replacement using tissue-engineered intervertebral discs in the canine cervical spine. Full Text Available The most common reason that adults in the United States see their physician is lower back or neck pain secondary to degenerative disc disease.
To date, approaches to treat degenerative disc disease are confined to purely mechanical devices designed to either eliminate or enable flexibility of the diseased motion segment. Tissue engineered intervertebral discs TE-IVDs have been proposed as an alternative approach and have shown promise in replacing native IVD in the rodent tail spine.
TE-IVD components were constructed using adult canine annulus fibrosis and nucleus pulposus cells seeded into collagen and alginate hydrogels, respectively. Seeded gels were formed into a single disc unit using molds designed from the geometry of the canine spine. Stably implanted TE-IVDs demonstrated significant retention of disc height and physiological hydration compared to discectomy control.
Both 4-week and week histological assessments demonstrated chondrocytic cells surrounded by proteoglycan-rich matrices in the NP and by fibrocartilaginous matrices in the AF portions of implanted TE-IVDs. Integration into host tissue was confirmed over 16 weeks without any signs of immune reaction. Despite the significant biomechanical demands of the beagle cervical spine, our stably implanted TE-IVDs maintained their position, structure and hydration as well as disc height over 16 weeks in vivo.
Heterogeneity of functional properties of Clone 66 murine breast cancer cells expressing various stem cell phenotypes. However, there is a lack of studies comparing the functions of CSCs isolated using different phenotypes in order to determine if CSCs are homogeneous or heterogeneous.
Cells with various stem cell phenotypes were isolated by sorting from Clone 66 murine breast cancer cells that grow orthotopically in immune intact syngeneic mice.
These populations were compared by in vitro functional assays for proliferation, growth, sphere and colony formation; and in vivo limiting dilution analysis of tumorigenesis. Differences in frequency and size of tumor spheres from these populations were observed. In vivo, fewer cells with a stem cell phenotype were needed for tumor formation than “non-stem” cells.
Whether the heterogeneity reflects soluble factor production remains to be determined. These data demonstrate that Clone 66 murine breast cancer cells that express stem cell phenotypes are heterogeneous and exhibit different functional properties, and this may also be the case for human breast cancer stem cells. Discrimination of meniscal cell phenotypes using gene expression profiles.
Full Text Available The lack of quantitative and objective metrics to assess cartilage and meniscus cell phenotypes contributes to the challenges in fibrocartilage tissue engineering. Although functional assessment of the final resulting tissue is essential, initial characterization of cell sources and quantitative description of their progression towards the natural, desired cell phenotype would provide an effective tool in optimizing cell -based tissue engineering strategies.
The purpose of this study was to identify quantifiable characteristics of meniscal cells and thereby find phenotypical markers that could effectively categorize cells based on their tissue of origin cartilage, inner, middle, and outer meniscus. Application Area: Analyzing cell proliferation and death.
It’s allowing us to design experiments we hadn’t considered before. I especially like how it comes with a networked computer that is very fast, allowing us to analyse results from our own slower computers really quickly. It’s a very intuitive system to figure out and we’ve even trained a high school student to operate it. Some of the consumables are a bit expensive, but most are available from other vendors.
Overall, we’re very happy with the machine and it’s so heavily used here we’re working on getting another. Great system for gaining better insight into cellular mechanics. Application Area: Migration assay in disease models. The software analysis was a little non-user friendly to figure out at the beginning, but there is a lot of functionality in setting up and analyzing the cell migration. Live-cell imaging gives more insights into cellular activity than just the numbers alone, so this tool is a great addition to our disease modeling systems.
Workhorse imaging! Application Area: Live imaging of drug response in neurons. We regularly are screening for neuro protective compounds in our iPSC model system where we can gain invalueable real-time data of response with images and graphs of 5 phenotypes of neurite changes.
We can set up the images and times ahead of time with analysis in real-time and come back days later with super graphs of data. It is a highly-valued component of our research work. Reliable, good images for cell monolayers. Application Area: 3D cultures. Sometimes we struggle with the analysis of 3D spheroid cultures as it is difficult to mask using the current software not the latest IncuCyte machine.
However, we still find it very useful for the multiple assays we run in the lab. We would like to have an improved analytic tool for 3D cultures spheroids, organoids that could be compatible with the machine we have.
The IncuCyte Zoom has become an invaluable tool in our lab. Application Area: Cell proliferation, migration and tumour-immune cell interactions. All this in a “track-and-trace” fashion. We are able to track the behaviour of individual cells, as well as collect data on large pools. With multiple lasers installed and the possibilities of using many different plate formats, there are few limitations to possible combinations.
Being able to determine the use of the camera in the way that is desirable for a particular experiment also leads to individual optimisation so that the IncuCyte can be used by many researchers throughout the entire research institute, tackling many different research topics and questions. Overall, the versatility of the instrument is great, and if needed can be expanded eg. The downsides are rather minimal, but need to be mentioned. First of all, there is a limited space inside, so only 6 plates can be tracked at a time during an experiment; sometimes one would wish this to be more.
The user-interface is alright, but could be a bit more user-friendly. A good training course is given once a machine will be installed on-site, which is very helpful, but I feel that it would be quite hard to learn yourself, without help from an experienced user.
Last, as always, the equipment and maintenance of an IncuCyte is a pricey business. Also the reagents provided by the company are not cheap. Luckily there is very good tech support in case of need – it normally doesn’t take long to get one of their experts where you need them. Strongly recommended for fluorescence-based analysis of dynamic biological processes.
Application Area: Virology, fluorescence reporter assays, immunology. Introductions for new lab members do not take more than 30 min. Experimental setups are easy and straight-forward. Running multiple up to 6 different experiments at the same time by different researchers is realistically possible.
We use it a lot for plaque formation – even with very slow viruses. Imaging runs can be 10 days or more to assess plaque growth velocity or abortive infection at very low MOI. Minor criticism: There are some non-solvable hot pixel issues at very high contrast with our IncuCyte.
High background in green channel with some cell media probably riboflavin-associated – can be erased with e. ImageJ background subtraction. I would love to have a blue fluorescence channel in addition – that would make the perfect machine. I think this is the best cell biology equipment I have used.
Application Area: Immune cell and neuronal cell assay. The live-cell analysis system did all the repetitive, tedious work like taking pictures every 2 hours for 2 weeks which is almost impossible for me to do by myself, which makes me really get into the details of what happened in the cells after the treatment.
I can monitor cells on my computer any time I like and I can make decisions based on live-cell conditions. The software created results and figures directly, saved me tons of time to do the analysis. With IncuCyte, I am more confident in my results and I have a better and more clear way way to represent my data.
Sometimes you do experiments in normal ways and the results are kind of ambiguous; then with IncuCyte you get much much more detail, you get a much more clear picture to resolve the uncertainty. Used for every project. Application Area: Cell cytotoxicity assays. Simple to use with great imaging analysis outputs. Application Area: Bioimaging core facility. However, most microscope systems give you 96 well movies that you have to then analyse to get the live cell data graphs.
The Incucyte is a high content live-cell screening machine that gives you 96 well kinetic graphs at the end of the experiment, that also has the movies for you to watch back if you want to. It’s about approaching the imaging experiment from what you want to measure and using the best tools to get there. The range of reagents are very good and it means that your experiment is just going to work. It takes just a couple of minutes to set up and experiment and the results are once you’ve set up the analysis pathway once there while the experiment is running.
We have loads of microscopes in the facility, but the IncuCyte continues to be one of the most popular. Really useful bit of kit and great after sales care!
Application Area: Analyzing cell growth in response to drugs or genetic manipulation. Support staff have always been extremely helpful and responsive and very efficient with call outs.
The IncuCyte lets you collect large amounts of data and compare multiple conditions very easily and really speeds up the science! Application Area: Compound efficacy on caspase activation and cell viability. I can monitor my experiments in real time.
If I have questions, whether it’s regarding protocols or analysis, a field application scientist from Sartorius will get back to me relatively quickly. Hopefully, this will change so I can access and analyze data from home.
Great tool needed in every cell culture lab. I’ve also found it very useful for cell killing assays including co-cultures of cells with activated T-cells.
Great instrument, some of the more technical uses of the machine require more teaching. The only thing that is hard with the machine is that it cannot handle 6 96 well plates and I hope that the machine can be upgraded in the future to be able to scan them without a break. Furthermore some technical support window for just routine data analysis would be nice.
User friendly. Application Area: Proliferation, chemotaxis, scartch wounds. For our current experiemnts, it is really easy to use and very user-friendly. I wish that it was easier to use different plates or create your own methods.
The chemotaxis plates from the company are a bit out of our budget. In general, IncyCyte Zoom is a great tool for capturing biology in real-time.
However, the assay window might not be robust enough to screen compounds. It really depends on the biology of the targets and the cell types. The real time imaging can pickup the dynamics but the biology might need to be verified by independent means if the assay window is too small. The best solution for long term live cell imaging. Application Area: Lice cells imaging assays. System offers the best features to perform lice cell imaging assays.
Ines S. Very nice instrument! Application Area: Cell viability. We analyze cell viability and cell death. Solid piece of kit. Application Area: Live cell imaging, proliferation time lapse, cell death modes, wound healing. We have it mounted in an incubator in our cell culture room and it is usually full. The ease of use, ability to track images from any computer, ability to generate a lot of data in a relatively small amount of time, and ability to analyze in many different modes has quickly secured its position as a user friendly workhorse of an instrument.
It became basic equipment almost instantaneously. Application Area: Basic research on molecular biology of cancer. A recent paper on effects of inhibitors on cell proliferation and migration was accepted because we could provide IncuCyte data. We have looked at nuclear location, cell toxicity, but also migration and apoptosis.
The wound maker works like a charm, although the users have to follow the instructions for this precious device. We recently changed the incubator, which now prevents too much condensation on the machine. Many colleagues are now also using our machine with succes for many different applications, showing its robustness. Very usefull and easy to use.
Application Area: Cell death assay, cell growth. Data analysis takes bit more time to learn. Very easy to use. Application Area: Pharmacology, cell biology, cancer. Machine very user-friendly, analysis tool not so much. The pictures are of good quality. Compatible with many different types of plates and slides. Drawbacks: The maximum magnification is only 20x, making it difficult to study e.
The built-in analysis tool is not always suitable to distinguish between different cells, and the plugin for single cell analysis is very expensive. Great tool for great results. Application Area: Microbiology. As a microbiologist, I follow intracellular fluorescent bacteria. Using Incucyte S3, I can assess and control 10 different conditions at the same time. It is a real time-saver and thus a saver of money.
Can’t imagine my project without this instrument. Application Area: Cancer cell migration, proliferation and invasion. Good results, unclear user practices.
Application Area: Cell toxicity assays. Application Area: For the growth of bacterial colonies. High quality results. You can check cells from home on weekend. Application Area: Analyze drug inhibition. No termination of the study and you can see the process. The only thing I dislike is that some cells grow on top of each other, which cannot be detected accurately by IncuCyte. Best multi-user real time imager for cell culture. Application Area: Real time quantification of apoptotic or necroptotic cell death.
It has showed increased speed, versatility and so far improved reliability. Our laboratory productivity performance is increased and we have already published data generated with our IncuCytes in several journals.
In summary, we strongly recommend this instrument and all of Sartorius’ supporting team behind it. Every lab needs one. Application Area: Analyze cell growth after drug treatments or gene editing. We get more reliable results in way less time than doing a manual approach. The machine was so busy our lab bought a second IncuCyte to accommodate the demand. The 96 well format allows for many replicates of different conditions all at the same time.
It’s also great to have images to look back on if you see an interesting phenotype. Would definitely recommend to anyone looking at cell growth. Great images, awesome scientific help with analysis. The only minor issue is that the analysis of the assays can sometimes be a little bit complicated. But I feel that customer service really covers that department and I’ve been lucky to get very good guidance from Essen Satorius Group.
We cannot live without this instrument! Application Area: Analysis of cellular responses to drug treatment: proliferation, motility, invasion, 3D growth. We utilize it in a “core facility” manner, so lots of labs share it. Our ZOOM is in almost constant use, running any number of applications. Almost everyone starts with proliferation studies, they are so much more informative in real-time than a static MTS assay.
We also employ the ImageLock plates for 2D motility or invasion in would healing-type assays. We use the ClearView plates to look at 3D motility and invasion. We grow 3D spheroids in wells to test chemotherapeutic drugs or combination therapies, in an attempt to model in vivo studies. There are several other modules that we don’t currently use, but we could unlock them with a phone call to Sartorius.
Flexibility is the key! This company goes beyond my wildest expectations for service and support. Many issues can be solved with a phone call and a WebEx and if not, a technician can be on-site within a day or so. Consistently provides quality data for proliferation, cell death, wound healing, etc. Application Area: Cancer Biology. The results are consistent, reliable, and easily reproducible.
The company provides great service and always responds promptly when we had an issue. I would recommend this to any lab looking at cellular assays. It is much easier and saves a ton of time compared to other assays or other applications requiring microscopy.
Great products – amazing and very cool. Application Area: Stem cells. Great instrument that’s easy to use! Application Area: Immune cell killing assays. Great instrument and I cannot think of working without it for my assays. Application Area: Cell growth assay. I immediately discarded all those assays in my mind when we bought the IncuCyte S3. It can count the number of cells and the confluency in the plates, at any time point, automatically and accurately, whether the whole cell population or subpopulations are labeled with different colors.
Additionally it may eventually replace many traditional assays like apoptosis, cell cycles. The instrument saves me a lot of time for my experiments and it has been amazing.
Great results, but needs constant servicing. Application Area: Cell death. We have had to service our Incucyte multiple times for various issues, which is the main drawback of the unit. Powerful and easy to use.
Application Area: Cell cycle, drug response. Can get reproducible data easily. Companion kits make designing experiments easy. Good customer service. Application Area: Tumor cell killing rates. I have liquid and tumor cells and use different immune cells including NK cells to look at the killing rate at different time points. Good results. Application Area: Metabolomics.
One can get reliable results. It’s very handy for spheroids, especially the 4x objective. Application Area: Growth kinetics, cytotoxicity of spheroids. User friendly interface, very easy to follow anaylsis. All of the application materials were easy to find and follow. Patrick was very helpful with the follow up questions. Very user friendly and versatile. Application Area: Confluency curves.
User care is great as a technician came to give an overview of the software and how to use. Application Area: Viral pathogenesis research.
It has potential to revolutionize the future of fluorescent microscopy and flow cytometry. Looking forward to the inclusion of more fluorescent or laser filters. I couldn’t run the experiments I have planned without it. Application Area: Cell killing. While CTG or other viability assays have been useful in the past, they have not been very accurate with our cell lines. The Incucyte has been a game changer as we design more experiments looking at cell killing.
At this point, I couldn’t run the experiments I have planned without it. Software is very user friendly and requires little to no training. Saves lots of time for long-term cell proliferation.
Application Area: Cell toxicity; cell proliferation. Single cell organoid growth with and without drugs for tracking over time. An automated solution for doing more than ever before with less effort. Not only that, I have been able to gather magnitudes more data and better quality data than before.
The Incucyte has removed a key bottleneck in our work and enabled us to do more than ever before. Application Area: Cell based assays. Application Area: Cell culture. Great results and gets nice data. Application Area: Neuron activity. It can be used in many fields and gets wonderful data. Can be used for multiple applications. Application Area: Neurite outgrowth. It is very intuitive and powerful with minimal hands-on time.
Incredible instrument with an awesome support team. Application Area: Proliferation Assays for drug screening. It is useful in a lot of research areas. The live-analysis helps in designing and planning of downstream experiments, while one experiment is still running.
With the green and red filters we use this instrument for a variety of bio-assays. I am especially impreesed by the customer care. All the application scientists are extremly helpful and very promt with thier responses. Many of them have come to help us beyond work hours if required. Great insight from the results on the target and effector cell interactions. High quality data and reproducibility can be very easily achieved. It’s value for money as multiple data sets are collected, and this can be presented in different ways as per the need.
It is fantastic! Application Area: Analyze cell proliferation, signal pathway response and gene edit efficient with reporter cell line. It looks simple and very easy to use. Initially I used the machine for cell proliferation curve generation and morphology observation which is the basic function.
Later on I tried to use it to detect fluorescence intensity response in my NFkB reporter cell line. It worked very well and was sensitive. The results are consistent and reliable. Recently I did an experiment for gene expression time response with different gene editing deliver system.
It saved a lot of time compared with using other methods. I am sure the machine has much more potential than I expect. I am also very satisfied with the service. Great machine! Application Area: Cell proliferation, cytotoxicity, migration, etc. Has a very easy interface, no need to have a lot of microscopy background to start an experiment.
Great for assay development! Application Area: Cell assay development. We use this equipment to monitor cell growth kinetics and chose ideal cell plating densities based on assay length, etc. Has revolutionized in the field of in-vitro cell biology.
Application Area: Cytotoxicity and internalization assay. It can do many more things such as cell proliferation, cytotoxicity, internalization chemotaxis, etc, without changing the media or disturbing the cells.
I love the improvements in the S3 software, imaging camera and automated objective lens control. Application Area: Live cell imaging. Image quality and quantitation of data is great. The software modules and reagents for specific applications are easy to use and always growing.
Very good. Application Area: Biotechnology product. Consistent results. Application Area: Cell proliferaction analysis. Nice to have one for your own lab! A great and reliable tool for high quality results!
Application Area: neuroscience, neurite growth, astrocyte adhesion and spreading, astrocyte proliferation, glutamate e. It allows a wide variety of analyses for cell phenotyping and drug testing. Great instrument, I can’t live without it! Application Area: neural cell based assay. Key instrument for determining dynamic range of our neuron outgrowth experiments. Application Area: Determine neurite changes after drug chemotherapy treatment. It allows us to pin-point a more precise range of damage occurring, with real-time graphing of data.
Saves us many weeks of optimization with only a single 3 day run on the IncuCyte S3. It radically changed the way we analyse cell proliferation. Application Area: Live-cell analysis platform to monitor cell proliferation, viability, apoptosis and a number of cell.
It is equipped with a powerful image analysis software which allows even beginners to get excellent results. The hardware is simple, robust and very easy to use. In our opinion, the IncuCyte has solved some classic drawbacks of time-lapse microscopy, since it has excellent optical stability, minimal sample disturbance, and consistent autofocus.
The possibility of integrating different objectives 4x, 10x and 20x and up to two fluorescence channels, in addition to the brightfield and phase contrast, allow it to perform most of the image-based cell assays with excellent resolution.
The IncuCyte is designed for aggregate cellular analysis it is not an image-based cytometer , but the introduction of the temporal component in the quantitative analysis is a parameter of inestimable importance. This powerful instrument can complement traditional cellular analytical approaches, both kinetic and end-point, and stands as the middle ground between low-resolution plate readers, time-lapse microscopes and high-content screening platforms.
The use of IncuCyte in daily research activities has allowed us to understand complex mechanisms of action of drugs that would have been difficult to extrapolate with classical approaches. In addition to this, it allowed us to speed up the phenotypic drug or loss of function screening via RNAi or CRISPR, as well as to make the cloning of modified cell lines more efficient. The IncuCyte must not be missing in a cell biology laboratory. Direct visualization and results. Application Area: cancer cell live observation – oxidative stress.
Great equipment. Application Area: Cell based assays, growth rate, dilution cloning,. The whole cell analysis can be done for vessels such as 6 well plates. The dilution cloning scan allows us to see one single cell.
Very useful. Great for cell functionality. Application Area: Cell-based assays for toxicity, drug delivery, and other cell based assays. It combines the instrument with reagents and image-based analysis into one system! Application Area: Single cell clone selection.
Multiple capacities of the incucyte. Application Area: follow live cells. Any trainee can use it after some user’s explanations. With this device, we are able to follow the growth of any cells and to have a morphologic view of the cells during time while leaving cells in their usual environment.
The software of image analysis included in this device allows to measure a lot of parameters such as the confluence of the culture or the neurites outgrowth. Many other possibilities exist with this device and the new version of the Incucyte Incucyte S 3 is even more performant. Super for quick and repeatable analysis. Application Area: lab demonstration.
The IncuCyte S3 is a crucial support for our cell-based assays. Application Area: Oncology and immuno-oncology. It integrates seamlessly into our screening funnel and has virtually replaced the traditional end-point proliferation assays. The IncuCyte S3 is a very powerful and easy-to-use tool at the same time, which I highly recommend to anyone involved in cell biology research.
Fantastic technology, no hands on perfectly autonomous. Application Area: Oncology research. We have been interacting with Essen biosciences on our expectations for the next generation of system, and there it is It is a very efficient equipment.
Application Area: In several live cell based monitoring and assays. I would recommend it highly. Easy and affordable, great to use! Application Area: Ovarian Cell Culture work and immune therapy. Great for migration and proliferation assays in the lab.
Would recommend to any cell culture user especially those interested in immune therapy. Our best Cell QC and monitoring system Application Area: Cell Quality Control. It allows many formats of cell vessels and the program is very intuitive! There are still a lot of applications we would like to test Thanks for this nice and helpful device!!!
Congratulations for being part of Sartorius Company Perfect for automated measurement of growth curves. Application Area: Measure cell proliferation. I can quickly set up an experiment to track cell proliferation and walk away while it collects data. It is very customizable and flexible – able to accommodate many different types of plates, and different imaging frequencies, and green and red channels in addition to phase images.
We obtained powerful results, more sensitive one and it is very save timing. Application Area: Analyze the ability of cell to proliferate well, with or without drugs causing DNA damage or breaks. We are working preferentially on fibroblast cells primary and SV40 transform cells. It allows us also to do a drug screening much more important. Of course there is some trouble that we can’t deal with for the moment, but we are sure that we can figure it out soon.
The increasing information that we got with the IncuCyte, leads us to ask ourself how to manage the data, and more precisely how to analyse and do statistics on it.
Engineer of Essen bioScience are very helpful about our question, but unfortunately whitout any regards it’s difficult to help us. The IncuCyte is very easy to use, the new software is much better than the old one and can be use easily even without anyone to help you. We are very satisfied of this equipment and have no regret to order it. Makes research so much easier! Application Area: Tissue culture and scratch wound assays. It allowed me to provide video data for my lab that is so much more interesting than just a graph!
It is so helpful to be able to run a bunch of assays on one plate as well. Really recommend!!! Excellent technology and worth the money. Application Area: Assess cell growth and function in controlled environment. In doing so, we are able to study cell growth and function in tightly controlled conditions without disturbing the cells. This has not been possible prior to the Incucyte technology, and we are very happy with our purchase. Great tool for good research.
Application Area: Proliferation, cell death, immune response. Very helpful for research, although an improvement of the optics switches and contrast of phase mask could improve the process. The best phase imaging system on the market. Application Area: Live cell imaging platform for labelled or unlabelled assays. The S3 has been brilliant in this regard, the software has been overhauled making everything much simpler and the camera is faster and has greater resolution than the ZOOM.
If I have any criticism its that the fluorescence imaging is not as sensitive as other systems but this is more than made up for by the excellent phase images.
Really Great. Application Area: Neurites outgrowth. I couldn’t do without it any more. The analysis module is really wonderful and very powerful!
The only way to improve it would be to add the blue light to easily take pictures of fixed cells. Outstanding performance. The system monitors proliferation by analyzing confluence and allows the separation of living and apoptotic cells by labeling reagents. Must have. Application Area: Functional Assays. The product itself is very easy to use. The software is very intuitive, in the beginning I was skeptical when the technicians from Essen told me this.
The results are nice and everything is analyzed by the machine itself and the power of this machine is the fact that you can let the experiment run as long as you want.
Makes many experiments a lot easier and safes work and time. Application Area: cell proliferation, cytotoxicity. We are also using it for spheroids and it works fine. Anyway, we are having some troubles with evaporation which leads to death of cells seeded in the edges of well plates, which is very annoying. It offers some basic statistic tools and allows to make good looking figures for presentations, for any more detailed analysis it is best to copy the data to a spreadsheet program like excel.
Instrument that should be a must in each lab. Application Area: Cell killing assay. Always getting my observation in real time and the analysis right away once the experiment was terminated. And everything with remote control. Application Area: Proliferation, cell death PI-uptake of cancer cells.
The IncuCyte system and software are very easy to use. After an intensive testing period and seminars on how to operate the equipment, I made the experience that handling the machine is easy and using the software is very intuitive. I recommend a “learning by doing” approach with first experiments, then all functions soon make sense. Organizing measurement schedules with colleagues works well due to the 24 hour display and the possibility to shift entire schedules until they fit.
The analysis will need a few rounds of “trial and error” – inaccuracies in detecting cells or staining are a trade-off for the convenient rapid high-throughput experiment and analysis, but can impact the result. Since images can be exported and analysed by more labor-intensive methods ImageJ , the output remains valuable, even if the semi-automatic analysis might not be able to gain the desired data.
The work load is very small and it is very convenient, how the software stores and analyses the data. I cannot comment on “After Sales Care”, since I didn’t use it. I am convinced that we will get value for the money, if the IncuCyte system is used regularly in the next few years. Great and easy. Application Area: Analyze cytotoxicity and apoptosis inducing activities.
Really useful in any lab. Good and easy to analyze results. Indispensable to our research. Additionally, saves a great deal of hands on time and allows for much more reproducible assays. Fits perfectly as part of a core facility platform. Wonderful product, a ton of results with just one experiment. Nice to have it for in-situ monitoring and for quantitative analysis. Application Area: Analyze anti-cancer efficacy of super molecular self-assembly.
And as we expected, it runs certain quantitative protocols efficiently, saved us a lot of time. I am satisfied with the performance of the machine. However, I was disappointed in the beginning. Because we received the new machine with problems of 4x objective. We had to send back the machine and the company promised to send us another new one with tested functions before it’s delivered.
Then we received a new one that had problem with the fluorescence module. We had to wait for the engineer sent a new part to replace the fluorescence module of the machine. The whole process was unpleasant since it wasted us quite a lot of time. I really want to ask did you test your machine before sending it out?
It’s intolerant that such problem happened twice. Very easy to use and fully automated. Application Area: Monitor spheroid growth and death with bright field image and fluorescent labeling. As long as the incubator in which the IncuCyte sits works, the live cell imaging can keep on going.
The onboard analysis is pretty nice too. Very useful instruments with greatly improved features. Application Area: 3D cell culture. The new software is actually much better than the old one, much more intuitive. I like Incucyte for its extensive recording and high throughput capabilities.
I was trying to adopt the spheroid assay to be used with higher concentration of Matrigel than recommended 2. For some reason, just opening the Incucyte would cause it to go out of focus on my spheroids so my images would have some pictures in focus and some not. Also in dense Matrigel spheroids not always settle at the very bottom of the plate and therefore the Incucyte has a hard time focusing on these ones.
Hardly a machine fault but still annoying for my purposes. It’s still nice to be able to use different objectives for different plates and be able to schedule different acquisition times for different vessels.
Grate improvement over the previous versions! User-friendly, great phase images. Application Area: Phenotypic Screening. High quality real-time results of cell culture. Application Area: Video microscopy analysis of cell culture behavior. Informs about adhesion, growth, migration, apoptosis, cell proliferation, neuritogenesis, angiogenesis, invasion and many other applications.
Great idea – but has reliability issues and underpowered image analysis software. Application Area: Live cell fluorescent imaging. When it worked, it worked fairly well. The image analysis software was adequate. The customer service was very helpful at bringing us up to speed with how to use the software, but it wasn’t all that powerful, and there were a few things that we thought would be straightforward that just weren’t possible. The images frequently drifted out of focus – so about half of our images were not useable – resulting in a tedious process of deleting every other photo from a movie.
I would have much rather just had the option of uploading images and movies to a server or the cloud, than having a bank of apparently unreliable hard drives. I liked the idea, and really wanted to love the instrument, but in the end I’m not sure I’ll buy another in my new lab. The Zoom is a cellular assay development scientist’s dream come true Another big plus is the “scan-on-demand” feature that enables additional plates to be imaged during unscheduled time.
Love that I can remote into the controller to adjust schedules, view plates, and perform analysis jobs anywhere I can go with my laptop internet connection required – it would be even nicer if there was a phone app The HD phase images are hard to beat and enable a number of very useful label-free analyses. Opportunities for improvement: 1 While the software interface definitely has the essentials and is relatively intuitive to learn and use, it could be vastly improved and modernized to enhance the user experience.
Overall, the Zoom is a great concept that quickly becomes a can’t-live-without-it lab instrument. Varied applications, reproducible results and great customer support! Application Area: Chemotaxis and Migration assays, proliferation assays. Most applications require an initial set up and then the IncuCyte Zoom takes care of the rest.
The wound maker is a big advantage for consistent and clean wounds in our adherent cells. The availability of multiple dish formats well, 24 well, 6 well also works out well when performing apoptosis assays and proliferation assays. The one disadvantage is the difficulty in changing objectives from 10x mostly preferred to 4x needed for whole well imaging. The instrument and scanning software is easy to use.
Remote access to the software allows multiple users to access data. However, the analysis part is not intuitive and requires multiple training sessions. There is a need to make this more user friendly. What the software lacks in ease of use is compensated by the many times Michael has stopped by to help us set up an experiment and analyse data.
We are literally fighting to sign up for the instrument. Overall, very happy with the instrument and its varied applications to cancer-related studies. Great results, very useful for our purpose. Application Area: neurons primary cell cultures and cell lines. Very useful instrument for cell growth and cytotoxicity studies but has some limitation compared to other high content imaging systems.
Application Area: Cytotoxicity and cell growth studies. The system is easy to set up and operate. The analysis of data produced by this system is a bit challenging and has a steep learning curve to fully utilize the full potential of analysis capabilities. This is a challenge for most mid to high content imaging systems due to the large amount of data generated. One significant limitation of the software is to identify multiple types of objects and produce multiple “masks” which would allow counting objects associated with other objects e.
This system also lacks light source and filters to detect blue fluorescence dyes such as DAPI and Hoescht. Great instrument, streamlines cell based research. Application Area: cell growth assays. I routinely run growth assays, wound healing, migration assays. It is easy to set up and I can monitor the progress live. Additionally, it can do on demand scans to measure confluency to eliminate variation in cell density sensitive experiments such as transient transfection.
Application Area: Live cell imaging – cell migration, invasion, apoptosis. The system and software are really easy to use and automatically captures high quality images in real-time.
Application Area: proliferation. Perfect to evaluate proliferation rates during assay development. Very reproducible results with a good experimental design and optimization. Application Area: well plate screening. The assays are based on detection of fluorescence. I really love the fact that one can do time courses and dose responses and get simultaneous morphological and functional information from each well in live cells!
Great instrument to have on hand! Application Area: cancer cell proliferation. The zoom has streamlined several key assays in lab. The ease of use both setup and analysis has impressed all researchers who have used the Zoom. The data produced has been viewed as high quality and essentially ready for presentations, posters, and publications. All in all, the Zoom has offered a highly streamlined approach for completing several key assays.
Application Area: T-cell killing of target cells Our old PI. Our new research P. Great high content imaging device! Customer service and sales representatives have been very helpful with any questions or problems we have had with the device. It has been a valuable tool to get high content imaging for meetings and presentations to better understand the activity of our molecules of interest.
Essential instrument in our lab. It has helped us tremendously and the results are excellent – very good quality images. Perfect tool for real-time monitoring of cell proliferation! Application Area: analyze cell proliferation rate and response to treatment.
Unique capabilities. Application Area: assessing cell behaviour over time. It is much easier to get different time-point results. Everyone should get one. Application Area: cell proliferation. In addition, the photos the thing takes are outstanding. Very handy instrument to get the results you want in an easy way but in low throughput.
Application Area: Analysis of cell growth, protein expression, morphology. Great results at a fast and high throughput capacity. Application Area: Imaging neuronal projections and live cell imaging. Great equipment with minor limitations. Application Area: Analyzing cellular phenotypes.
The quality of the images taken is decent but the capability to image sub-cellular components at higher magnification is lacking. One equipment takes over an entire incubator seems to be not so cost-effective. After all, it is a great tool for phenotypic study. Further, there are many secondary experiments that can be done at shorter, more frequent time scales. Application Area: neurite outgrowth and cytotoxicity. High efficiency.
Live cell video is excellent. We can monitor the growth of neuron and analysis the parameters of neurite and compared with the treatment group and non-treatment growth.
Good results so far. Application Area: To analyze cells. The results are good for a values publication which is an essential criterion.