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. 2013 Jun;16(6):779-83.

Co-culture of Mouse Embryonic Stem Cells with Sertoli Cells Promote in vitro Generation of Germ Cells

Mohammad Miryounesi  1 , Karim Nayernia, Mahdi Dianatpour, Fatemeh Mansouri, Mohammad Hossein Modarressi
Affiliations

Co-culture of Mouse Embryonic Stem Cells with Sertoli Cells Promote in vitro Generation of Germ Cells

Mohammad Miryounesi et al. Iran J Basic Med Sci. 2013 Jun.

Abstract

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 Stra8 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 (Stra8), meiotic (Dazl and Sycp3) and post-meiotic (Prm1) genes were evaluated at different differentiation stages (+7, +12 and +18 days of culture).

Results: In the first group, expressions of meiotic and post-meiotic genes started 12 and 18 days after induction with retinoic acid, respectively. In the second group, 7 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.

Keywords: Co-culture; Differentiation; Embryonic stem cell; In vitro derived germ cells; Sertoli cell.

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Figures

Figure 1
Figure 1
Co-culture of mESCs (arrows) colonies and Sertoli cells
Figure 2a
Figure 2a
Diagram of GFP expressed mESCs sorted by FACS. b and c: Sorted cells showed high expression of GFP
Figure 3
Figure 3
Expression of meiotic (Sycp3 and Dazl) and postmeiotic (Prm1) genes in two groups of culture: Group 1: culture of mESCs on gelatin with RA treatment,Group 2: co-cultured of mESCs with Sertoli cell feeder without RA induction
See this image and copyright information in PMC

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