Induction of Ram Bone Marrow Mesenchymal Stem Cells into Germ Cell Lineage using Transforming Growth Factor‐β Superfamily Growth Factors

Abstract

Several studies have proposed that in vitro generation of germ cells (GCs) from stem cells can be considered a future option for infertility treatment. Mesenchymal stem cells (MSCs) have the capability to differentiate into male GCs with the use of inducers such as retinoic acid. Transforming growth factor-beta 1 (TGFb1) has been shown to play important roles in male fertility and spermatogenesis. Bone morphogenic protein 4 (BMP4) and BMP8b are also involved in the derivation of primordial GCs (PGCs) from epiblast cells. Therefore, this study aims to determine whether TGFb1, BMP4 and BMP8b can initiate transdifferentiation of MSCs into GCs in vitro and to determine the type of changes that occur in the expression of GC-specific markers. In this study, we have divided passage-3 ram bone marrow (BM)-MSCs into three main groups (BMP4, BMP8b and TGFb1) which were separately treated with 10 ng/ml TGFb1, 100 ng/ml BMP4 and 100 ng/ml BMP8b for a period of 21 days. We have evaluated the ability of these groups to differentiate into GCs by assessing expressions of GC-specific markers with reverse transcription PCR (RT-PCR), quantitative RT-PCR (qRT-PCR), immunocytochemistry, morphological changes and alkaline phosphatase (ALP) activity. Our results showed that BMP4 and BMP8b induced PGCs properties in some BM-MSCs and TGFb1 formed spermatogonial stem cells (SSCs) and spermatogonia-like cells in BM-MSCs culture. The important results of this study provide the basis for additional studies to determine the exact mechanism of GCs differentiation and possibly solve the problem of infertility.

Overexpression of STRA8, BOULE, and DAZL Genes Promotes Goat Bone Marrow-Derived Mesenchymal Stem Cells In Vitro Transdifferentiation Toward Putative Male Germ Cells

“…These results demonstrate that a population of BMSCs might also show germ cell characteristics without treatment, which has previously been confirmed in human, ram, and mouse MSCs. 8,10,11 In addition, these studies showed that there was a temporal and spatial correlation between OCT4 and MVH expression patterns. Generally, the downregulation Figure 7.…”

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

“…A number of in vitro studies confirmed that MSCs have the capacity to differentiate into GCs (13, 18, 20–24). Transplantation studies on the effects of MSCs on reconstruction of testicular germinal epithelium in infertile male animals, showed a number of promising results.…”

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

“…As previously mentioned, under appropriate conditions, MSCs can differentiate into male and female GCs in vitro (Drusenheimer et al 2007;Ghasemzadeh-Hasankolaei et al 2014b;Hua et al 2009a, b;Huang et al 2010;Nayernia et al 2006;Qiu et al 2013). Moreover, it has been indicated that MSCs possess a population of germ-like cells that express GC-specific markers (Drusenheimer et al 2007;Johnson et al 2005;Woodbury et al 2002).…”

“…A number of research groups have focused on the differentiation capacity of MSCs into the GCs. Their reports have shown that under appropriate conditions, MSCs from different sources such as fetal and adult BM, umbilical cord Wharton's jelly and lungs can form male and female GCs in vitro (Drusenheimer et al 2007;Ghasemzadeh-Hasankolaei et al 2014b;Hua et al 2009a, b;Huang et al 2010;Nayernia et al 2006;Qiu et al 2013). These papers have also shown that untreated MSCs exhibit some characteristics of GCs such as expression of GCspecific genes.…”