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Background and aims: Infertility is one of the most common problems among couples. Generation of male germ cells from adult stem cells is a current promising priority of researchers. This study aimed to investigate the potential of human umbilical cord mesenchymal stem cells (hUMSCs) on the expression of male germ cell markers after isolating by this method. Methods: The hUMSCs was incubated with retinoic acid, testosterone, and conditioned medium (prepared from testicular cell cultures of 7-day-old mice) during 3 days. The bands were visualized and densitometry was accomplished using LI-COR Biosciences software. Results: The high expression levels of C-KIT, DAZL, PIWIL2, and DDX4 in mRNA and protein levels were observed in treated hUMSCs. Conclusion: Results of reverse transcription polymerase chain reaction (RT-PCR) and western blotting showed that method of isolation had no adverse effects on differentiation potential of hUMSCs. [ABSTRACT FROM AUTHOR]