Investigation of expression of miR-134 in differentiation of Amniotic Membrane-derived Mesenchymal Cells to Neural-like cells

Background and Aim : Amniotic membrane mesenchymal stem cells (MSCs) are pluripotential mesenchymal stem cells that differentiate from different tissues. Mesenchymal stem cells obtained from amniotic membrane are valuable and easy to access which do not need to surgery. They can also be differentiation to any type of cells such as osteocyte, adipocyte, and neural cells. Amniotic membrane is the area of placenta which originates from extraembryonic tissues. Amniotic membrane mesenchymal stem cells have self-renewal, angiogenesis, differentiation, and flexibility features and we can differentiate these cells into neural cells and use to treat diseases and neural disorders. miRs have an important role in differentiation of neural cells and increase in neural cells, one of which is miR-134 that we investigated in this study.Methods : In this study, we derived mesenchymal stem cells from human amniotic membrane; to confirm their mesenchymal feature, surface markers of mesenchymal stem cells were determined by flowcytometry. In order to examine their ability to be differentiate, the mesenchymal stem cells differentiate into osteocyte and adipocyte lineages. Then, they differentiate to neural-like cells by adding the neural differentiation environment containing bFGF, EGF, butyric acid, Indomethacin, Ascorbic acid and their features were investigated. After extraction of RNA and cDNA synthesis, the expression of miR-134 was determined by the Real Time PCR technique.Results : Through analysis of flow cytometry was specified that CD90,44,105 markers are positive while CD34,45 markers were negative. The accumulation of adipocyte vesicles in cells which were exposed to adipocyte medium is the indicator of their differentiation to adipocyte cells. Also, cells had showed calcium deposits when they were placed in osteocyte medium. after treating the amniotic membrane mesenchymal stem cells with the neural differentiation environment, we observed change in the morphology of mesenchymal stem cells which were differentiated to neural-like cells. The level of miR-134 expression in neural-like mesenchymal stem cells increased 15 time more than witness cells.Conclusion : The study of neural-like cells is very important. Therefore, according to the results, it seems that using the differentiation cocktail amniotic membrane mesenchymal stem cells were differentiate into neural- like cells, which can be applied in treatment of neurological diseases and disorders such as Alzheimer, Huntington, Parkinson and the stroke in future.

Keywords:

mesenchymal stem cells , differentiation , neural- like cells , amniotic membrane , miR-134