Transcriptional profiling of early differentiation of primary human mesenchymal stem cells into chondrocytes
Ontology highlight
ABSTRACT:
INSTRUMENT(S): Illumina Genome Analyzer IIx
ORGANISM(S): Homo sapiens
SUBMITTER: Thomas Schwarzl
PROVIDER: E-MTAB-10476 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:The treatment of bone defects caused by infection, trauma or neoplasms remains a clinical challenge. Autologous bone transplantation is limited by availability, donor site morbidity and surgical risk factors. This has given rise to stromal/stem-cell based therapy. Bone marrow derived stromal cells (BMSCs) have been studied to a large extent and show high regenerative potential but their use is limited by availability, donor site morbidity and the relatively low cell yield as they represent only <0.1% of cell harvested from bone marrow aspirate. At the same time, they are the closest mesenchymal stromal cells for bone tissue engineering given their tissue origin and, unlike other mesenchymal stromal cells, can support the formation of hematopoietic marrow. Adipose tissue derived stromal cells (ASCs) as part of the stromal vascular fraction of adipose tissue can as well undergo osteogenic differentiation but can be additionally isolated in a sufficient quantity from lipoaspirate after liposuction of abundant subcutaneous fat tissue. Here, it has been shown that there are no major differences in regard to proliferation or differentiation capacity of ASCs derived from subcutaneous fat of different anatomical regions. It has been shown that BMSCs are more prone to senescence during expansion and passage than ASCs and that ageing impacts proliferative capabilities of BMSCs more than that of ASCs while it has also been reported that osteogenic differentiation capacity is least impacted by age. Multiple studies have compared the characteristics of these two mesenchymal stromal cells in regard to bone tissue engineering in vitro. Most studies point to inferior extracellular matrix mineralization and lower expression of key osteogenic transcription markers like Runx2 in osteogenic differentiated ASCs compared to BMSCs. On the other hand, a study by Rath et al. found contrary results using particular culturing conditions like 3D bioglass scaffolds. An intraindividual comparison of human MSCs of three donors cultured on decellularized porcine bone confirmed superior osteogenic capacity of BMSCs compared to ASCs. In contrast to BMSCs, ASCs were not able to induce heterogenic ossification in a mouse model. In a sheep tibia defect model application of BMSCs resulted in a significantly higher amount of newly formed bone tissue. Importantly, Osteogenic differentiated ASCs do not support the formation of a hematopoietic marrow. Proteomics enables large-scale analysis of proteins present in a cell type and can be used to identify differentially regulated key proteins in a comparative approach. A comparative proteomic analysis of BMSCs and ASCs by Roche et al. in 2009 identified 556 proteins with 78% of these not being differentially regulated between these two cell populations, regarded as high similarity. Another comparative proteomic study of 2016 by Jeon et al. found 90 differentially regulated proteins out of 3000 total identified proteins. Both studies do not specify a number of different tissue donors and in part using cell lines. Looking for differences upon osteogenic differentiation, transcriptomic comparison of osteogenic differentiated porcine ASCs and BMSCs has been performed, resulting in 21 differentially expressed genes after 21 days of osteogenic culture conditions. Still, it remains unanswered, which are the key distinctive features of osteogenic differentiated ASCs and BMSCs at protein level that might help address the abovementioned weaknesses of ASCs in bone tissue engineering/regeneration for translational research. To overcome this need, an intraindividual comparative DIA based proteomic analysis of osteogenic differentiated human BMSC and ASCs was performed in this study.
2021-06-09 | PXD015223 | Pride
Project description:Transcriptional activation of cultured mouse astrocytes in response to stimulation with CCM (complete cytokine mix: TNF-alpha, IL1-beta and IFN-gamma) at 4hr and 16hr time points.
2005-12-01 | E-MEXP-321 | biostudies-arrayexpress
Project description:The main goal of the present work was to use an in vitro adipogenic-differentiation mode, with mesenchymal stem-cells (MSC) from human bone-marrow, to study whole-transcriptomic differential gene-expression when such cells are induced to differentiate into adipocytes in absence or presence of oleuropein. See details below. This work is complementary to a previously published one (Casado-Daz et al, 2016): Casado-DÃaz, A., Anter, J., Müller, S., Winter, P., Quesada-Gómez, J. M. and Dorado, G. (2016): Transcriptomic analyses of adipocyte differentiation from human mesenchymal stromal-cells (MSC). Journal of Cellular Physiology 9999: 1-14. DOI: 10.1002/jcp.25472. It corresponds to EMBL-EBI ArrayExpress accession E-MTAB-4884.
2018-12-31 | E-MTAB-5211 | biostudies-arrayexpress
Project description:Growth & differentiation mechanisms of Bone marrow derived mesenchymal stem cells.
2007-08-25 | E-TABM-318 | biostudies-arrayexpress
Project description:To see the effect of zoledronate on the osteogenic differentiation of human mesenchymal stem cells. Cells were first cultured in normal media until enough cells were obtained. Then the cells were differentiated into ostepbalst under the effect of low concentration of zoledronate.
2020-01-16 | E-MTAB-7459 | biostudies-arrayexpress
Project description:Wounds, especially non healing wounds are characterised by elevated tissue lactate concentrations. Lactate is known for being able to stimulate collagen synthesis and vessel growth. Lately it has been shown that lactate, in vivo, plays an important role in homing of stem cells. With this work we aimed to show the influence of lactate on the gene expressionprofil of human mesenchymal stem cells (hMSC). hMSCs were obtained from bone marrow and characterised with fluorescence-activated cell sorting (FACS) analysis. Subsequently the hMSCs were treated with either 0, 5, 10 and 15 mM lactate (pH 7,4) for 24 hours. RNA Isolation from stimulated hMSCs and controls was performed. The Microarray analysis was performed using Affymetrix HuGene 1.0 ST Gene Chip. Selected targets were subsequently analysed using quantitative real time PCR (RTq-PCR). We were able to show that lactate in moderate concentrations of 5 respectively 10 mM leads to an anti-imflammatory, anti-apoptotic but growth and proliferation promoting gene expression after 24 h. In contrast, high latate concentrations of 15 mM leads to the opposed effect, namely promoting inflammation and apoptosis. Hypoxia induced genes did not not show any significant regulation. Contrary to expectation, we were not able to show any significant regulation of glycolysis associate candiadtes. We were able to show that lactate alters gene expression but does not change the cell phenotype, which might be helpful for further investigations of new treatment strategies for chronic non-healing wounds as well as tumor-therapy and neuronal plasticity. Timecourse experiment with hMSCs treated with 5, 10, 15 mM lactic acid for 1, 3, 7 days. hMSCs without lactate served as controls *** CEL files lost due to hard disk crash
2013-01-09 | E-GEOD-40205 | biostudies-arrayexpress
Project description:Cells resident in tissues must be resilient to the physical demands of their surroundings. Our current understanding of cellular mechano-signalling is largely based on static systems, but these models do not reproduce the dynamic nature of living tissue. Here, we examined the time-resolved response of primary human mesenchymal stem cells (hMSCs) to periods of cyclic tensile strain (CTS). We observed parallels between morphological changes following low-intensity strain (1 hour, 4% CTS at 1 Hz) and responses to increased substrate stiffness. However, as the strain regime was intensified (CTS at ≥ 2 Hz), we characterised a broad, structured and reversible protein-level response, even as transcription was apparently shut down. Regulation of the linker of nucleo- and cytoskeleton (LINC) complex proteins, and specifically of SUN domain-containing protein 2 (SUN2), was found to decouple mechano-transmission within the cell and hence isolate the nucleus from cellular deformation.
2019-04-24 | PXD012949 | Pride
Project description:The repertoire of growth factors determines the biological engagement of human mesenchymal stromal cells (hMSCs) in processes such as immunomodulation and tissue repair. Hypoxia is a strong modulator of the secretome and well known stimuli to increase the secretion of pro-angiogenic molecules. In this manuscript, we employed a high throughput screening assay on an hMSCs cell line in order to identify small molecules that mimic hypoxia. Importantly, we show that the effect of these small molecules was cell type/species dependent, but we identified phenanthroline as a robust hit in several cell types. We show that phenanthroline induces high expression of hypoxia-target genes in hMSCs when compared with deferoxamine (DFO) (a.k.a desferrioxamine B, a known hypoxia mimic) and hypoxia incubator (2% O2). Interestingly, our microarray and proteomics analysis show that only phenanthroline induced high expression and secretion of another angiogenic cytokine, interleukin-8, suggesting that the mechanism of phenanthroline-induced hypoxia is distinct from DFO and hypoxia and involves the activation of other signaling pathways. We showed that phenanthroline alone was sufficient to induce blood vessel formation in a Matrigel plug assay in vivo paving the way to its application in ischeamic-related diseases.
2018-10-04 | E-MTAB-7209 | biostudies-arrayexpress
Project description:Adipogenesis is a physiological process required for fat-tissue development, mainly involved in regulating the organism energetic-state. Abnormal distribution-changes and dysfunctions in such tissue are associated to different pathologies. Adipocytes are generated from progenitor cells, via a complex differentiating process not yet well understood. Therefore, we investigated differential mRNA and miRNA expression patterns of human mesenchymal stromal-cells (MSC) induced and not induced to differentiate into adipocytes by next (second)-generation sequencing. A total of 2,866 differentially-expressed genes (101 encoding miRNA) were identified, with 705 (46 encoding miRNA) being upregulated in adipogenesis. They were related to different pathways, including PPARG, lipid, carbohydrate and energy metabolism, redox, membrane-organelle biosynthesis and endocrine system. Downregulated genes were related to extracellular matrix and cell migration, proliferation and differentiation. Analyses of mRNA-miRNA interaction showed that repressed miRNA-encoding genes can act downregulating PPARG-related genes; mostly the PPARG activator (PPARGC1A). Induced miRNA-encoding genes regulate downregulated genes related to TGFB1. These results shed new light to understand adipose-tissue differentiation and physiology, increasing our knowledge about pathologies like obesity, type-2 diabetes and osteoporosis.
2016-08-31 | E-MTAB-4884 | biostudies-arrayexpress
Project description:The response of osteoprogenitors to calcium (Ca2+) is of primary interest for both normal bone homeostasis and the clinical field of bone regeneration. The latter makes use of calcium phosphate-based bone void fillers to heal bone defects, but it is currently not known how Ca2+ released from these ceramic materials influences cells in situ. Here, we have created an in vitro environment with high extracellular Ca2+ concentration and investigated the response of human bone marrow-derived mesenchymal stromal cells (hMSCs) to it. Ca2+ enhanced proliferation and morphological changes in hMSCs. Moreover, the expression of osteogenic genes is highly increased. A 3-fold up-regulation of BMP-2 is observed after only 6 h and pharmaceutical interference with a number of proteins involved in Ca2+ sensing showed that not the calcium sensing receptor, but rather type L voltage-gated calcium channels are involved in mediating the signaling pathway between extracellular Ca2+ and BMP-2 expression. MEK1/2 activity is essential for the effect of Ca2+ and using microarray analysis, we have identified c-Fos as an early Ca2+ response gene. We have demonstrated that hMSC osteogenesis can be induced via extracellular Ca2+, a simple and economic way of priming hMSCs for bone tissue engineering applications. For more information check: https://cbit.maastrichtuniversity.nl/
2018-10-06 | E-MTAB-7222 | biostudies-arrayexpress