Possible Involvement of Pigment Epithelium Derived Factor (PEDF) in the Proliferation and Differentiation of Mouse Spermatogonial Cells in Vitro - Israeli Fertility Association Conference-Ayala 2018, May 7-8, Dan Panorama Hotel, Tel-Aviv

Noy Bagdadi ¹ Eitan Lunenfeld ² Mahmoud Huleihel ³

¹The Shraga Segal Dep. of Microbiology, Immunology and Genetics, The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, Ben-Gurion University of the Negev
²Dep. OB/GYN, Soroka Medical Center, The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, Ben-Gurion University of the Negev
³The Shraga Segal Dep. of Microbiology, Immunology and Genetics, The Center of Advanced Research and Education in Reproduction (CARER), Faculty of Health Sciences, The National Institute for Biotechnology in the Negev, Ben-Gurion University of the Negev

Introduction: Pigment epithelium derived factor (PEDF) is released as a soluble, monomeric glycoprotein, encoded by SERPINE-1 gene. Spermatogenesis occurs in the seminiferous tubules of the testis, which are avascular. Recently, it has been shown that cultures of human testicular peritubular cells constitutively secrete PEDF; and was suggested that PEDF may prevent vascularization of human seminiferous tubules. Furthermore, we have shown that PEDF is produced by mouse Sertoli, Leydig and peritubular cells (unpublished data).

Aim: To examine the possible involvement of PEDF in development of mouse spermatogenesis in vitro.

Materials and methods: Cells were enzymatically isolated from the seminiferous tubules of 7-day-old immature mice. Cells were cultured in metheylcellulose (as a 3D in vitro culture system) and KSR with/without PEDF (1-100 ng/ml). After 4-6 weeks of culture, the developed cells and colonies were collected and evaluated by immunofluorescence staining for pre-meiotic, meiotic and post-meiotic cells by using antibodies to markers specific for each type of cells.

Results: Our results showed development of colonies (proliferation) from isolated tubular cells in our system after 4 weeks of culture. Addition of PEDF to the isolated cells increased the percentage of the premeiotic cells (VASA) and meiotic cells (BOULE), but did not affect ACROSIN-positive cells compared to control after 4 weeks of culture. This effect of PEDF was in a dose-dependent manner.

Conclusions: This is the first study to show a direct effect of PEDF on the proliferation and differentiation of spermatogonial cells. These results may suggest the possible involvement of PEDF in the development of spermatogenesis under normal conditions.