Objective: From the distant past, many studies have shown the effects of zinc on male infertility. Zinc is a well-known essential trace element that plays a unique role in polymeric organization of macromolecules like DNA and RNA, DNA transcription, expression of steroid receptors, proteins synthesis, and cell division. Zinc is involved in the activation and maintenance of the germinal epithelium of semineferous tubules and able to affect germ cells for more deviation and differentiation. This is the first report about the effects of zinc ion on expression of male germ cells features in mesenchymal stem cells. Materials and Methods: Passage-3 male sheep marrow- derived mesenchymal stem cells treated with 0.14 μg/ml zinc sulfate (ZnSO4) for 21 days. Expression of male germ cells features was evaluated by assessment of germ cells common features include expression of germ cell-specific markers like VASA, PIWIL2, OCT4, beta1 INTEGRIN, DAZL (by RT-PCR, qRT-PCR) and PGP 9.5 (by immunocytichemistry), morphological characteristics and changes of the cells alkaline phosphatase activity. Results: No morphological changes were seen by treatment. Although VASA, PIWIL2, OCT4 and beta1 INTEGRIN were expressed in control group, but the expression of VASA and beta1 INTEGRIN were upregulated and PIWIL2 and OCT4 were downregulated. DAZL and PGP 9.5 weren’t expressed and alkaline phosphatase activity did not change after zinc treatment. Conclusion: These results show that although ZnSO4 doesn’t induce germ cell differentiation in mesenchymal stem cells but upregulation of VASA and beta1 INTEGRIN and OCT4 downregulation, suggests that maybe one of the mechanisms by which zinc ion can increase male fertility is help to regulate the expression of some specific genes in male germ cells of the testis and their differentiation process.