The BioInvestigationIndex (BII)

Title:

Human mesenchymal stem cells induced to differentiate by treatment with the NSAID indomethacin and/or other drugs. [original title: miR-335 regulates cell migration and differentiation in human mesenchymal stem cells]

Organism(s):

Homo sapiens

Description:

Total RNA was collected, and miRNA expression was compared from mesenchymal stem cells (MSCs) induced to differentiate along adipogenic or osteogenic pathways or transduced with miR-335. Controls: uninduced MSCs, skin fibroblasts, and MSCs transduced with a control vector. [original 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 remodeling, 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.]

Design(s):

transcription profiling by array

Experimental factor(s):

drug 1 conc 2 recorded
0.01 uM 0.1 uM
NSAID conc 1 recorded
60 uM
NSAID 2 recorded
indometacin none
condition 6 recorded
adipogenic induction overexpressing miR335 osteogenic induction control vector undifferentiated not specified
tissue 2 recorded
bone marrow skin
drug 2 conc 1 recorded
0.5 mM
drug 1 2 recorded
dexamethasone none
drug 2 2 recorded
3-isobutyl-1-methylxanthine none
chemical 2 conc 1 recorded
0.2 mM
chemical 1 2 recorded
none glycerol 2-phosphate
chemical 1 conc 1 recorded
10 mM
cell type 2 recorded
mesenchymal stem cell skin fibroblast
chemical 2 2 recorded
none ascorbic acid

Publication(s):

Tomé M, López-Romero P, Albo C, Sepúlveda JC, Fernández-Gutiérrez B, Dopazo A, Bernad A, González MA.
miR-335 orchestrates cell proliferation, migration and differentiation in human mesenchymal stem cells. PubMed:21164520

Sample attribute(s):

Material Type 2 recorded
total_RNA unknown
condition 5 recorded
adipogenic induction overexpressing miR335 osteogenic induction control vector undifferentiated
Cell line type 1 recorded
primary cell
tissue 2 recorded
bone marrow skin
Label 1 recorded
Cy3
cell type 2 recorded
mesenchymal stem cell skin fibroblast
organism 1 recorded
Homo sapiens