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

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.

Fertility preservation practices in pediatric and adolescent cancer patients undergoing HSCT in Europe: a population-based survey

“…Prepubertal boys present a significant challenge to FP because the only option that can be offered at the moment is cryopreservation of spermatogonial stem cells. 29 This procedure has emerged only in recent years and was not allowed in all participating countries at the time the EBMT survey was carried out; in fact, only 11 centers offered this method as an FP option. Spermatogonial stem cell cryopreservation represents a highly experimental technique and has only been proven to be effective in a mouse model; thus, its acceptance by professionals and parents is low, 30 in line with the low prevalence of its use reported in this survey.…”

Effect of semen washing methods on diminishing the transmission of viral infections in artificial reproductive technology

“…All of the infertility factors in men have not been identified yet and more comprehensive studies are required (1). Approximately, 15% of male infertility is related to genital and urinary tract infectious agents (2,3).…”

Xeno- and transgene-free reprogramming of mesenchymal stem cells toward the cells expressing neural markers using exosome treatments

“…A major obstacle to the reprogramming of somatic stem cells to commit to the cell types from a different germ layer is their epigenetic memory in the form of DNA methylation, limiting the potency of the cells. Previous studies showed that epigenetic modifications with 5-Bromo-2'deoxyuridine (BrdU) [1] or decitabine (5-aza-2'deoxycytidine) [19], increased the potency of MSCs. Decitabine induces DNA demethylation by blocking DNA methyltransferase and prevent differentiation of neural progenitors [20].…”