Study of Expression of miR-124 in Neural-like cells Differentiated of Mesenchymal Stem Cell Derived from human Amniotic membrane (hAMSCs)

Background and Aim : The amniotic membrane is the innermost of the three layers forming the fetal membranes and a valuable source of mesenchymal stem cells. The mesenchymal stem cells isolated from amniotic membrane have unique features including Self-Renewal ،developing and repairing damaged tissues and failure to stimulate the immune system. they are easily available and do not cause any controversial issues in their recovery and applications. Since the nervous system after injury who limited capacity to repair itself, For this reason in this study, amniotic membrane-derived mesenchymal stem cells were differentiated into nerve cells to investigate their differentiative features.Methods : After the isolation of stem cells from amniotic membrane in order to approve their mesenchymal features, their surface marker were evaluated by flow cytometry. Then, they were differentiated to osteocyte and adipocyte to assess their differentiative potential. Finally, the cells were exposed to the neural differentiation medium including Retinoic acid and ascorbic acid and at last was studied morphology of cells. The expression of miR-124 were studied with Real-Time PCR method.Results : Flow cytometry analysis showed that the expression of CD73, CD44, CD29, CD105 and CD73 markers was positive and the expression of CD34 and CD45 markers was negative. Calcium accumulation and adipose vesicles respectively showed the differentiation of mesenchymal cells to osteocyte and adipocyte. Morphology of mesenchymal cells was spindle shaped that was changed under the influence of neural differentiation medium and tails similar to axon and dendrites were observed in them. Real-Time PCR results showed the expression of miR-124 increase in the neural like-cells compared to the control group.Conclusion : According the results, since miR-124 was identified as one of the up-regulated miRNAs in the mammalian brain, it was identified as a neurogenic initiator.thus increasing of miR-124 indicate the differentiation of mesenchymal stem cells toward neural cells.

Keywords:

Mesenchymal Stem Cell , Amniotic membrane , Neural- like Cell , miR-124