Male and female rat bone marrow-derived mesenchymal stem cells are different in terms of the expression of germ cell specific genes

Abstract

Recent studies have shown that mesenchymal stem cells (MSCs), under appropriate conditions, can differentiate into cell types including germ cells (GCs). These studies also show that MSCs without any induction express some GC-specific genes innately. Moreover, one report suggests that female MSCs have a greater tendency to differentiate into female instead of male GCs. Therefore, for the first time, this study attempts to assay and determine the differences between the expression levels of some important GC-specific genes (Stra8, Vasa, Dazl, Stella, Piwil2, Oct4, Fragilis, Rnf17 and c-Kit) in male and female bone marrow (BM)-MSCs of rats. BM sampling of the rate was performed by a newly established method. We cultured rat BM samples, then characterized male and female MSCs according to their adhesion onto the culture dish, their differentiation potential into bone, cartilage and fat cells, and phenotype analysis by flow cytometry. The expression of GC-specific genes and their expression levels were evaluated with reverse transcription polymerase chain reaction (RT-PCR) and real-time RT-PCR. Our results showed that Dazl and Rnf17 did not express in the cells. The majority of examined genes, except Piwil2, expressed at almost the same levels in male and female MSCs. Piwil2 had higher expression in male MSCs which was probably related to the more prominent role of Piwil2 in the male GC development process. Male BM-MSCs appeared more prone to differentiate into male rather than female GCs. Additional research should be performed to determine the exact role of different genes in the male and female GC development process.

Transplantation of Autologous Bone Marrow Mesenchymal Stem Cells into the Testes of Infertile Male Rats and New Germ Cell Formation

“…BM sampling and isolation of BM-MSCs were performed according to our previously published paper (31). Isolated cells were characterized using the criteria proposed by the International Society for Cellular Therapy defined as: adhesion onto the bottom of the culture dish, expression of MSC CD markers, lack of expression of other cell lineage CD markers, and the capacity of differentiation into osteocytes, adipocytes and chondrocytes (32).…”

“…BM sampling and isolation of BM-MSCs were performed according to a new protocol reported in our recently published paper (31). Briefly, after induction of anesthesia in the rats (with an intraperitoneal injection of 60 mg/kg ketamine and 6 mg/kg xylazine) and dorsal recumbency, the knee region was shaved carefully and disinfected using 70% ethanol and povidone iodine.…”

“…For induction of differentiation into bone, fat and cartilage, P3 BM isolated cells were treated with osteogenic, adipogenic and chondrogenic media which was the same culture media used in our previous study (31) for 21 days. Generation of osteocytes, adipocytes and chondrocytes was confirmed by specific staining (alizarin red for osteogenic, oil red O for adipogenic and toluidine blue for chondrogenic differentiation), reverse transcription polymerase chain reaction (RT-PCR) and detection of the expressions of bone, fat and cartilage specific genes in the cells after 21 days treatment.…”

Erkek Rat Testislerinde Busulfan Kaynaklı Hasara Karşı Vitamin E, Melatonin, L-karnitin ve Atorvastatin’in Koruyucu Etkisi: Gen Ekspresyonunun Değerlendirilmesi

“…Mean efficiency values (E) for each gene were also determined from the amplification profiles of individual samples using the same software [7] . The following formula was applied to determine the relative gene expression in tissue testes of treated rat compared to the control group [18,19] .…”

In vitro differentiation of bovine bone marrow‐derived mesenchymal stem cells into male germ cells by exposure to exogenous bioactive factors

“…Among adult stem cells, MSC may be suitable candidates for in vitro gamete derivation and subsequent use in cell transplantation due to: (i) plasticity that is not limited to mesodermal derivatives, (ii) availability of abundant sources of tissues for isolation in several animal species including bovine, (iii) high proliferative potential, (iv) simple and inexpensive isolation from foetal and adult somatic tissues and (v) high potential for cell therapy including autologous or allogeneic transplantation due to reduced intrinsic teratogenic formation and immune rejection. Recent studies have explored the GC‐specific gene expression profile in male and female rat MSC (Ghasemzadeh‐Hasankolaei, Eslaminejad, Batavani, & Ghasemzadeh‐Hasankolaei, ) and evaluated the transdifferentiation potential of MSC derived from mice amniotic membrane (Afsartala et al., ), sheep bone marrow (Ghasemzadeh‐Hasankolaei, Eslaminejad, Batavani, & Sedighi‐Gilani, ), and human umbilical cord Wharton′s jelly (Huang et al., ) and bone marrow (Hua, Yu, et al., ). These studies have generated important evidence that supports the potential capacity of MSC for differentiation into male GC lineage.…”