Homo sapiens
Ontology highlight
ABSTRACT:
PROVIDER: PRJNA120755 | ENA |
REPOSITORIES: ENA
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Project description:Human mesenchymal stem cells (hMSC) have an extensive potential for clinical applications in cell therapy. However, very little is known of the specific molecular regulatory mechanisms that control the therapeutical properties of these cells. We aimed to identify microRNAs (miRNAs) that could be involved in controlling the transition between the self-renewing (undifferentiated) and the reparative (differentiated) phenotypes of hMSCs. MicroRNA microarrays were used to identify miRNAs that are upregulated in undifferentiated hMSCs. For that, we compared the miRNA expression profiles of undifferentiated bone marrow-derived hMSCs with the same primary cell lines after 9 days of in vitro adipogenic or osteogenic induction. We also compared the miRNA expression profiles of undifferentiated hMSCs with skin fibroblasts (a mesenchymal cell lineage with a more restricted differentiation potential). These experiments allowed us to identify miR-335 as the only miRNA downregulated upon MSC differentiation as well as in MSCs in comparison with skin fibroblasts. Gene expression microarrays were used to identify genes that are downregulated in hMSCs overexpressing miR-335. We compared the miRNA expression profiles of hMSCs transduced with a lentiviral vector encoding miR-335 with MSCs transduced with a control lentiviral vector. Our results suggest miR-335 downregulation could be one of the triggers for the initiation of MSCs activities involved in tissue repair and remodelling, including cell migration and differentiation. We compared the miRNA expression profiles of undifferentiated bone marrow-derived hMSCs with the same primary cell lines after 9 days of adipogenic or osteogenic induction, as well as with skin fibroblasts. A total of four independent samples were used for each condition. For the adipogenic/osteogenic vs. undifferentiated MSC comparison, the RNA samples were pooled (two independent samples/pool) before labeling. We also compared the miRNA expression profiles of hMSCs transduced with the lentiviral vector pLV-EmGFP-MIRN335 with MSCs transduced with the control vector pLV-EmGFP-Mock. For the gene expression microarrays, a total of three independent samples were used for each condition.
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Project description:Corneal diseases are a leading contributor of vision impairment and their treatment remains challenging. It’s known that enhancement of the stemness of corneal epithelial stem cells (CECs) plays a pivotal role in effective treatments. In this study, we evaluated the effects of extracellular vesicles from human adipose-derived mesenchymal stem cells (AdMSC-EVs) on CECs. Firstly, colony formation assays showed that the colony-forming efficiency (CFE) of CECs significantly increased in the presence of AdMSC-EVs. Then we demonstrated that AdMSC-EVs accelerated migration of CECs in a scratch assay, whereas the proliferation of CECs was suppressed by AdMSC-EVs in cell proliferation assay. RNA Sequencing (RNA-Seq) analysis of CECs implied that AdMSC-EVs maintained the stemness, as well as improved epithelial-mesenchymal transition (EMT). Furthermore, having identified the six top microRNAs (miRNAs) of AdMSC-EVs, CEC transfection with those miRNA mimics, indicated that miR-25, miR-191, and miR-335 were the most effective molecules for improving stemness and EMT. Taken together, our findings indicated that AdMSC-EVs can enhance stemness and EMT of CECs, and AdMSC-EV effects are exerted by the contained miRNAs.
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