Role of spermatogonial stem cells extract in transdifferentiation of 5-Aza-2′-deoxycytidine-treated bone marrow mesenchymal stem cells into germ-like cells

Role of spermatogonial stem cells extract in transdifferentiation of 5-Aza-2′-deoxycytidine-treated bone marrow mesenchymal stem cells into germ-like cells Kharizinejad E1, Minaee Zanganeh B1, Khanlarkhani N1, Mortezaee K1, Rastegar T1, Baazm M2, Abolhassani F1, Sajjadi SM3, Hajian M1, Aliakbari F1, Barbarestani M1. Abstract As one of the induced pluripotent stem cells (iPSCs) methods, spermatogonial stem cells (SSCS ) extract is considered as new approach in stem cell therapy of infertility. 5-aza-2'-deoxycytidine (5-aza-dC) inhibits methyltransferase enzyme, and induces gene reprogramming; herein, the effects of SSCS extract incubation in 5-aza-dC-treated bone marrow mesenchymal stem cells (BMMSCs) has been surveyed. BMMSCs were isolated from femurs of three to four weeks old male NMRI mice, and the cells at passage three were treated with 2 µM 5-aza-dC for 72 hours. SSCs were isolated, cultured, and harvested at passage three to collect SSCS extract; BMMSCs were then incubated with SSCS extract in the three time periods: 72 hours, one week and two weeks. There were five groups: control, sham, extract, 5-aza-dC and extract-5-aza-dC. After one week of incubation, flow cytometry and real-time polymerase chain reaction (PCR) exhibited high levels of expression for β1- and α6-integrins and promyelocytic leukaemia zinc finger (PLZF) in extract and extract-5-aza-dC groups (P < 0.05 vs. control and 5-aza-dC), and cells in these two groups had two forms of morphology, round and fusiform, similar to germ-like cells. 5-aza-dC had no significant effects during the three time periods of evaluation. These data disclose the effectiveness of SSCs extract incubation in transdifferentiation of BMMSCs into germ-like cells; this strategy could introduce a new approach for treatment of male infertility in clinic. Microsc. Res. Tech. 79:365-373, 2016. © 2016 Wiley Periodicals, Inc. KEYWORDS: 5-aza-2′-deoxycytidine; spermatogonial stem cells extract; transdifferentiation