A gene expression profile analysis of the differentiation of muscle-derived stem cells into smooth muscle cells from sheep.

ABSTRACT: OBJECTIVE:To analyze gene expression profiles analysis during the differentiation of sheep muscle-derived stem cells (MDSCs) into smooth muscle cells (SMCs) in sheep. METHODS:A modified preplate technique was employed to isolation of the MDSCs from sheep. The MDSCs were subjected to flow cytometry analysis targeting CD44, CD31, CD45, CD14, and CD49f and were treated with TGF-?1 at a concentration of 10 ng/ml for ten days. The expression levels of smooth muscle ?-actin (?-SMA) and calponin after treatment with TGF-?1 were determined by western blotting and immunofluorescence staining. A microarray analysis was performed to screen for differentially expressed genes (DEGs) during MDSC differentiation using total RNA extracted from MDSCs and SMCs generated from MDSCs. Molecule Annotation System (MAS) 3.0, which employs KEGG (Kyoto Encyclopedia of Genes and Genomes) and GO (Gene Ontology) Consortium annotations, was used to identify global biological trends in the gene expression data. RESULTS:The expression levels of the SMC-specific contractile proteins ?-SMA and calponin were dramatically increased after treatment with TGF-?1. Immunofluorescece staining showed that the TGF-?1-treated MDSCs were positive for ?-SMA. We identified 486 genes that were differentially expressed between the MDSCs and TGF-?1-treated MDSCs. 260 of which were up-regulated and 226 of which were down-regulated. Twenty-one genes exhibited a greater than ten-fold change, 13 of which were up-regulated and 8 of which were down-regulated. TGF-?1 treatment up-regulated both the SMAD and MAPK signaling pathways during the differentiation of these sheep cells. The PPAR and Wnt signaling pathways were also found to be involved in the differentiation process. CONCLUSIONS:TGF-?1 can successfully induce the differentiation of sheep MDSCs into SMCs. For the first time, we analyzed the gene expression profiles associated with this differentiation process, and the results showed that both the SMAD and MAPK signaling pathways are involved. This study indicated that multiple signaling networks coordinate the development and differentiation of MDSCs into SMCs.

SUBMITTER: Tang X 

PROVIDER: S-EPMC5934578 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature