Endometrial Stem Cells Differentiation Into Neural Cells by Ly294002 Small Molecule



Kouchesfehani HM¹ ; Ebrahimibarough S² ; Ai J² ; Anbar H¹ Show Affiliations
Source: Koomesh Published:2016

Abstract

Introduction: Endometrial stem cells (EnSCs) were identified for the first time in 2004. These cells are capable of extensive self-renewal and have the potency to differentiate into chondrocyte, osteocyte, adipocyte, neuron and oligodendrocyte. PI3K/Akt signaling has been implicated in multiple cellular and organ functions, including differentiation, survival and cell death and its inhibition leads to cell differentiation. The purpose of this study was to investigate the differentiation of endometrial stem cells into neural cells by inhibition of PI3K/Akt pathway using small molecule Ly294002. Materials and Methods: Endometrial tissues were treated enzymatically and segregated cells were cultured in DMEM/F12 with 10% FBS. The flow cytometry analysis was perfomed for CD105, CD90, CD146, CD31 and CD34 at the third passage. Then the neurogenic differentiation was evaluated at the third passage, 21 days after induction with differentiation media. Immunocytochemistry and Real-time PCR were performed to investigate the expression of specific neural stem cells markers. Results: The flow cytometry analysis showed that EnSCs were positive for CD90, CD105 and CD146 and negative for CD31 and CD34. Immunocytochemistry showed that the expression of nestin, NF and Chat neuronal markers in the cells treated with small molecule Ly294002. Real-time PCR also indicated expression of NF and Chat neuronal markers at the mRNA level. Conclusion: According to the findings of this study it can be concluded that the EnSCs have neural differentiation potency in the suitable differentiation milieu. Ly294002 small molecules by inhibiting PI3K / Akt pathway possibly can prevent cell proliferation and induce cell differentiation. © 2016, Semnan University of Medical Sciences. All rights reserved.