Effect of Zinc Ions on Differentiation of Bone Marrow-Derived Mesenchymal Stem Cells to Male Germ Cells and Some Germ Cell-Specific Gene Expression in Rams

Abstract

This is the first report to describe the effects of zinc (Zn) ions on the expression of germ cell (GC) genes from bone marrow-derived mesenchymal stem cells (BM-MSCs). Zn plays an important role in germinal epithelium maintenance, testosterone secretion, differentiation of GCs, and spermatogenesis. In addition, several studies have suggested that MSCs have the potential for differentiation into numerous cells types, including male GCs. In this study, we have treated passage-3 ram BM-MSCs with 0.14 μg/ml Zn sulfate (ZnSO₄) for a period of 21 days with the intent to determine whether Zn treatment can stimulate MSCs to differentiate into male GCs in vitro. We also sought to determine the type of changes seen in MSCs by Zn treatment. Differentiation into male GCs was evaluated by the assessment of expressions of the following GC-specific markers: VASA, PIWIL2, OCT4, beta1 INTEGRIN (ITG b1), DAZL (by reverse transcription polymerase chain reaction (RT-PCR) and quantitative RT-PCR), and PGP 9.5 (by immunocytochemistry). Also studied were morphological characteristics and changes in alkaline phosphatase activity. Interestingly, Zn upregulated the expressions of VASA and ITG b1 but downregulated PIWIL2 and OCT4. DAZL and PGP 9.5 were not expressed in the treatment group. According to our results, Zn ions did not stimulate BM-MSCs to transdifferentiate into male GCs; however, it changed the expression of GC genes in BM-MSCs. It can be concluded that a possible mechanism by which Zn ions can increase male fertility is by regulation of the expression of testis GC-specific genes in the differentiation process and spermatogenesis.

Enhancement of osteogenic differentiation of rat adipose tissue-derived mesenchymal stem cells by zinc sulphate under electromagnetic field via the PKA, ERK1/2 and Wnt/β-catenin signaling pathways

“…After 15–20 min, alizarin red 40 mM (pH 4.1) was added to each well. The plates were incubated at room temperature for 20 min with shaken and then they were washed 2–3 times with PBS shaken for 5 min to reduce nonspecific staining [5, 36]. …”

“…Chondrogenic differentiation medium composed of high-glucose DMEM with 10% FBS, 100 U/mL penicillin, 100 μg/mL streptomycin, 1.3 mM L-ascorbic 2-phosphate, 0.01 mM dexamethasone and ITS+1 Liquid Media Supplement (100x) was used for 14 days. At the end of chondrogenesis, cells were fixed with 4% (v/v) paraformaldehyde for 30 min, washed with PBS for three times and proteoglycans were stained with 0.1% toluidine blue [36]. …”

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

“…The RT-PCR process was performed according to former reports (33, 34). Briefly, TRIzol reagent (Invitrogen, Paisley, UK) was used to extract total RNA from the cells according to the manufacturer’s instructions.…”

Emerging Methods to Generate Artificial Germ Cells from Stem Cells1

“…1). Some evidence has shown that germ-like cells can be generated from somatic stem cells derived from mouse [30,31], sheep [32,33], or human [34][35][36] mesenchymal stem cells; fetal mouse skin [37,38] as well as porcine skin [39][40][41][42] or muscle [42,43] stem cells; and human amniotic fluid stem cells [44,45]. Combined, these studies provide examples in which somatic stem cells differentiated into male or female germ cells in culture.…”