Isolation of Mesenchymal Stem Cells from Human Adipose Tissue and Comparison of miRNA-16 and miRNA-125b Expression before and after Differentiation into Adipocytes and Liver Cells
Abstract
Introduction: The hallmark of stem cells is their ability to produce numerous differentiated cells. The use of complementary compositions such as ketorolac and triamcinolone can improve the biological processes. In addition, expression profile of microRNAs (miRNAs) in the commitment of mesenchymal stem cells (MSCs) is unknown. Present study aimed to evaluate miRNAs expression levels under different induction conditions. Methods: MSCs were isolated, expanded and directed using adipogenesis medium or medium supplemented with ketorolac, and also subjected to hepatogenic differentiation either with or without triamcinolone acetonide. Periodic acid-Schiff (PAS) staining, Oil red O staining, adiponectin and albumin secretion were evaluated. MiR-125b and miR-16 family expression level was assessed using qRT-PCR in four induced groups as compared to non-induced cells. Results: Multilineage potency of ADSCs was assessed using osteogenic induction. MiR-16 and miR-15 over-expressed significantly from adipose derived stem cells to adipocytes during 28 days (p < 0.01). Upregulation of miR-125b and miR16 was achieved during hepatogenesis (p < 0.05). Ketorolac stimulated upregulation of miR16/15b in the adipogenic induced stem cells. Interestingly, triamcinolone caused upregulation of miR-15b and miR-195 in hepatic commitment. Discussion: The results support the potential use of ADSCs in different applications. It also emphasized on involvement of the miR-16 and miR-125b in differentiation and yielded results under different induction conditions. It is suggested that miRNAs or complementary compounds can stimulate stem cells proliferation and differentiation; or applied as a tracking marker which provide new insights for future approaches in biomedicine.