Busulfan Treatment of Immature Mice Decreased the Amount of Spermatogonial Cells but Increased the Expression of Testicular Growth Fctors by Somatic Cells - Israeli Fertility Association Conference-Ayala 2017, May 8-9, 2017

Ali Abu Madigim ¹ Ronnie Solomon ¹ Maram Abofoul-Azab ¹ Eitan Lunenfeld ² Mahmoud Huleihel ¹

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

Introduction: Spermatogenic cells at different stages express specific markers. The proliferation and differentiation of spermatogenic cells are under hormonal and testicular growth factors control such as SCF,GDNF and CSF-1. All these factors affect proliferation of spermatogonial cells that express receptors for these factors. Busulfan is an alkylating agent used in cancer therapy that significantly affects male fertility mainly through apoptosis of the dividing cells – spermatogonial cells.

Aim: To evaluate the effect of busulfan on the amount of spermatogonial cells and on the levels of testicular growth factors.

Materials and methods: Immature mice were divided into control and busulfan groups. Mice were injected with a single dose of busulfan (45 mg/kg) or DMSO. Mice were sacrificed after 10 days. Cells were enzymatically isolated from the seminiferous tubules and count and stained for markers specific for spermatogonial cells (pre-meiotic), meiotic and post-meiotic cells, by immunofluorescence staining using specific antibodies or examined for RNA expression by qPCR analysis. Cells were also examined for growth factor expression by qPCR analysis.

Results: Our results showed a significant decrease in the total number of cells isolated from the tubules of the BU-treated mice compared to control, and also in the subpopulations of the spermatogonial cells. However, a significant increase was detected in the growth factors levels in cells isolated from BU-treated mice compared to control.

Conclusions: Our results show for the first time a direct effect of busulfan in the different subpopulation of spermatogonial cells. We suggest a compensatory mechanism by increasing the production of their growth factors.