Dedifferentiation of Granulosa Cells into Induced Pluripotential Stem Cells by Exposure to the Embryonic Stem Cell Extract (Pages: 0-0)

Kharazi E *, Rohani L , Talaei T , Vojdani Z , Mostafavipour Z , Tabeie Z ,


Objective: Epigenetic reprogramming of terminally differentiated cell can modify somatic cells to a pluripotential state. There are several approaches that induce pluripotency in somatic cells. Exposure the cells with the embryonic stem cell extract is an easy way, and some investigations were done on fibroblast cell line. However, its efficiency was low. So, the objective of this study was to increase the number of reprogrammed granulosa cell as a full differentiated cell into pluripotential state. Materials and Methods: human granulosa cells were cultured in medium containing azacytidine and trichostatine. The cells were exposed to mouse embryonic stem cells extract. The granulosa cells cocultured with mouse embryonic fibroblast in the presence of leukemia inhibitory factor. to assay the reprogrammed cells, alkaline phosphatase test and also immonocytochemistery were performed for OCT4, SOX2 and Nanog antibodies after 72 hours. Results: The results indicated that granulosa cells showed the alkaline phosphatase activity. They also express OCT4, SOX2 and nanog after exposure to the embryonic stem cells extract. Conclusion: It seems that the extract could induce dedifferentiation in granulosa cells. The previous research that was done on fibroblast cell line revealed the extract could lead the cells to express OCT4 but not nanog. We found that granolusa cells, as full differentiated somatic cells also can express the stem cell markers. It seems that the inhibitors of the methyl transferase and histone deacetylase could wipe the epigenetic markers and prepare the cells for reprogramming by administration of the extract.