Spermatogonial Stem Cell Subpopulation Survive Busulfan Treatment in Immature Mice - Israeli Fertility Association Conference-Ayala 2018, May 7-8, Dan Panorama Hotel, Tel-Aviv

Ali AbuMadigem ¹ Ronnie Solomon ² Eitan Lunenfeld ³ Mahmoud Huleihel ⁴

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
³Dep. 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
⁴The 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: Busulfan (BU) is an aggressive chemotherapy that used in bone-marrow transplantation and as anti-cancer treatment. Chemotherapy may lead to male infertility by killing the proliferating spermatogonial cells (SPGCs). However, whether subtypes of SPGCs survive the chemotherapy treatment has not yet studied.

Aims: To examine the effect of BU-treatment on SPGC markers of immature mice.

Materials and Methods: 7-day-old mice were injected intraperitoneally with BU (45mg/kg) or DMSO (control). Mice were sacrificed after 10-days (a point in time with nadir effect of BU). Cells from seminiferous tubules (STs) were enzymatically isolated and stained by immunofluorescence stained or identified by flow cytometry (FACS) using specific markers for SPGCs, and their RNA expression levels were evaluated by qPCR.

Results: A significant decrease in the number of SPGCs that express c-kit, g-csfr, thy1, sall4 and vasa with no change in the number of cells that express α-6-intgrin were examined in cells isolated from STs of BU-treated mice compared to the control group. Also, gene expression of the SPGC markers sall4, plzf, cdh1, thy1, vasa and gfr-α were decreased, but no change in the gene expression of the markers c-kit, α-6-intgrin and cd9 compared to control group.

Conclusions: We have shown for the first time that BU distinctly affected subpopulations of SPGCs from immature mice. Our results show that some of the SPGCs are resistance and survive BU treatment in immature mice. These results may deepen our understanding of chemotherapy effect on male infertility, and may open new therapeutic strategies for male fertility preservation mainly for prepubertal cancer patient boys.