Expression pattern of microRNA-16 family members’ during induction processes to mesodermal and non-mesodermal lineages with ketorolac and triamcinolone acetonide.

The role of miR-16/15b/195 and complementary compounds in the commitment of mesenchymal stem cells (MSCs) to different lineages is unknown. The aim of this work was to investigate the effect of ketorolac and triamcinolone acetonide on adipogenic and hepatogenic processes, respectively. Also, miR-16/15 and miR-195 expression levels were evaluated under different induction conditions.

MSCs were isolated, expanded and directed using adipogenesis medium or medium supplemented with ketorolac, and also subjected to hepatogenic differentiation using a cocktail of hepatocyte growth factor (HGF) and Oncostatin M (OSM) either with or without triamcinolone acetonide. Periodic acid-Schiff (PAS) staining, Oil red O staining, albumin and adiponectin protein secretion were evaluated. MiR-16 family expression level was assessed using qRT-PCR in four induced groups as compared to non-induced cells.

The expression levels of miR-16 and miR-15 increased significantly from adipose derived stem cells to adipocytes (p<0.01). Positive stimulatory effects of ketorolac and triamcinolone on adipogenic and hepatogenic induction, respectively, were observed. Ketorolac stimulated upregulation of miR-16/15b in the adipogenic induced stem cells. Interestingly, triamcinolone caused upregulation of miR-15b and miR-195 in hepatic commitment.

Two highly effective stimulators were identified. Ketorolac is similar to indomethacin and affects adipogenic differentiation. Triamcinolone is involved in hepatogenic commitment of stem cells like differentiation process of macrophages, adipocytes and osteocytes. The alteration of miR-16 family members’ expression indicates that this family may play a possible role in directing stem cell fate.