Dataset Information

Gene expression profiling: identification of gene expression in human MSC chondrogenic differentiation.

ABSTRACT: Understanding the mechanisms that govern cell fate will lead to the development of techniques for the induction of human mesenchymal stem cell differentiation into desired cell outcomes and the production of an autologous source of tissue for regenerative medicine. Here, we demonstrate that stem cells derived from adult bone marrow grown with 3D pellets take on characteristics similar to human cartilage. The NFAT signaling pathway is primarily linked to cell differentiation and influences chondrogenic differentiation. Based on our previous results that alterations in the expression of the NFATc1 gene affect chondrogenesis, we screened a microarray and identified 29 genes with altered expression, including 13 up-regulated (fold change ? 2) and 16 down-regulated (fold change ? 2) genes, compared with the control group. We then used RT-PCR to validate the chip data. Gene ontology and pathway analyses were performed on these altered genes. We found that these altered genes function in the complement and coagulation cascades, metabolism, biosynthesis, transcriptional regulation, proteolysis, and intracellular signaling pathways, such as the cytoplasmic calcineurin-dependent signaling pathway, the cyclin-dependent kinase inhibitor 2C signaling pathway, the MAPK signaling pathway, and the insulin signaling pathway. Our study suggests that these pathways may play important roles in chondrogenesis, which could be useful in the design of biomaterials.

SUBMITTER: Gong M

PROVIDER: S-EPMC6291687 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Gene expression profiling: identification of gene expression in human MSC chondrogenic differentiation.

Gong Ming M Liang Tangzhao T Zhang Hao H Chen Shaochu S Hu Yawei Y Zhou Jianhua J Zhang Xuan X Zhang Wang W Geng Xiaojing X Zou Xuenong X

American journal of translational research 20181115 11

Understanding the mechanisms that govern cell fate will lead to the development of techniques for the induction of human mesenchymal stem cell differentiation into desired cell outcomes and the production of an autologous source of tissue for regenerative medicine. Here, we demonstrate that stem cells derived from adult bone marrow grown with 3D pellets take on characteristics similar to human cartilage. The NFAT signaling pathway is primarily linked to cell differentiation and influences chondr ...[more]

PMID: 30662607

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Project description:Tissue engineering strategies often aim to direct tissue formation by mimicking conditions progenitor cells experience within native tissues. For example, to create cartilage in vitro, researchers often aim to replicate the biochemical and mechanical milieu cells experience during cartilage formation in the developing limb bud. This includes stimulating progenitors with TGF-?1/3, culturing under hypoxic conditions, and regulating mechanosensory pathways using biomaterials that control substrate stiffness and/or cell shape. However, as progenitors differentiate down the chondrogenic lineage, the pathways that regulate their responses to mechanotransduction, hypoxia and TGF-? may not act independently, but rather also impact one another, influencing overall cell response. Here, to better understand hypoxia's influence on mechanoregulatory-mediated chondrogenesis, we cultured human marrow stromal/mesenchymal stem cells (hMSC) on soft (0.167?kPa) or stiff (49.6?kPa) polyacrylamide hydrogels in chondrogenic medium containing TGF-?3. We then compared cell morphology, phosphorylated myosin light chain 2 staining, and chondrogenic gene expression under normoxic and hypoxic conditions, in the presence and absence of pharmacological inhibition of cytoskeletal tension. We show that on soft compared to stiff substrates, hypoxia prompts hMSC to adopt more spread morphologies, assemble in compact mesenchymal condensation-like colonies, and upregulate NCAM expression, and that inhibition of cytoskeletal tension negates hypoxia-mediated upregulation of molecular markers of chondrogenesis, including COL2A1 and SOX9. Taken together, our findings support a role for hypoxia in regulating hMSC morphology, cytoskeletal tension and chondrogenesis, and that hypoxia's effects are modulated, at least in part, by mechanosensitive pathways. Our insights into how hypoxia impacts mechanoregulation of chondrogenesis in hMSC may improve strategies to develop tissue engineered cartilage. STATEMENT OF SIGNIFICANCE: Cartilage tissue engineering strategies often aim to drive progenitor cell differentiation by replicating the local environment of the native tissue, including by regulating oxygen concentration and mechanical stiffness. However, the pathways that regulate cellular responses to mechanotransduction and hypoxia may not act independently, but rather also impact one another. Here, we show that on soft, but not stiff surfaces, hypoxia impacts human MSC (hMSC) morphology and colony formation, and inhibition of cytoskeletal tension negates the hypoxia-mediated upregulation of molecular markers of chondrogenesis. These observations suggest that hypoxia's effects during hMSC chondrogenesis are modulated, at least in part, by mechanosensitive pathways, and may impact strategies to develop scaffolds for cartilage tissue engineering, as hypoxia's chondrogenic effects may be enhanced on soft materials.

Dataset Information

Gene expression profiling: identification of gene expression in human MSC chondrogenic differentiation.

Publications

Gene expression profiling: identification of gene expression in human MSC chondrogenic differentiation.

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