Level of miR-101a and miR-107 in Human Adipose Mesenchymal Stem Cells Committed to Insulin-producing Cells.

Abstract

Mesenchymal stem cells have the fundamental ability to differentiate into multiple cells such as osteoblasts, neural cells, and insulin-producing cells. MicroRNAs (miRNAs) are single-strand and small non-coding RNAs involved in stem cells orientation into mature cells. There is no comprehensive data about the dynamic of distinct miRNAs during the differentiation of mesenchymal cells from adipose tissue into insulin-producing cells. In this study, we first differentiated adipose-derived mesenchymal stem cells into insulin-producing cells by a three-stepwise protocol. Differentiation capacity was confirmed by the dithizone staining method and hormone (insulin and C peptide) release analysis via electrochemiluminescence technique. In the final phase, the expression of hsa-miR-101a and hsa-miR-107 and two pancreatic genes, sex-determining region Y-box (SOX) 6 and neuronal differentiation 1 (NeuroD1) were examined during the differentiation procedure on days 0, 7, 14, 21, and 28 after induction, by using real-time PCR assay. The level of C-peptide and insulin were also measured at the end of the experiment. Dithizone staining showed trans-differentiation of adipose-derived mesenchymal stem cells into pancreatic β cells evidenced with red-to-brown appearance compared to the control group, indicating the potency to insulin production. These features were at maximum levels 28 days after cell differentiation. Real-time PCR revealed the increase of NeuroD1 and reduction of SOX6 during differentiation of stem cells toward insulin-producing cells (P <0.05). Both miR-101a and miR-107 showed prominent expression at day 28 (P <0.05). Changes in the expression of miR-101a and miR-107coincided with alteration of NeuroD1 and SOX6 that could affect mesenchymal stem cells commitment toward insulin-like beta cells.