Mohammad Miryounesi - Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
Karim Nayernia - GENEOCELL, Advanced Molecular & Cellular Technologies, Montreal, Canada
Mahdi Dianatpour - Department of Medical Genetics, School of Medicine, Shiraz University of Medical Sciences, Shiraz, Iran
Fatemeh Mansouri - Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran
Mohammad Hossein Modarressi* - Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

  Objective(s): Sertoli cells support in vivo germ cell production; but, its exact mechanism has not been well understood. The present study was designed to analyze the effect of Sertoli cells in differentiation of mouse embryonic stem cells (mESCs) to germ cells.   Materials and Methods: A fusion construct composed of a Stra۸ gene promoter and the coding region of enhanced green fluorescence protein was produced to select differentiated mESCs. To analyze sertoli cells’ effect in differentiation process, mESCs were separated into two groups: the first group was cultured on gelatin with retinoic acid treatment and the second group was co-cultured with sertoli cell feeder without retinoic acid induction. Expressions of pre-meiotic (Stra۸), meiotic (Dazl and Sycp۳) and post-meiotic (Prm۱) genes were evaluated at different differentiation stages (+۷, +۱۲ and +۱۸ days of culture). Results: In the first group, expressions of meiotic and post-meiotic genes started ۱۲ and ۱۸ days after induction with retinoic acid, respectively. In the second group, ۷ days after co-culturing with Sertoli cells, expression of meiotic and post-meiotic genes was observed. Conclusion: These results show that differentiation process to germ cells is supported by Sertoli cells. Our findings provide a novel effective approach for generation of germ cell in vitro and studying the interaction of germ cells with their niche.

Co-culture Differentiation Embryonic stem cell In vitro derived germ cells Sertoli cell