Xenogeneic spermatogenesis following transplantation of human spermatogonial stem cells to recipient mouse testes

Abstract

Exogenesis (cross-species) germ cell transplantation provides an opportunity to investigate fundamental aspects of spermatogenesis. This technique establishes a functional assay for testicular stem cells and allows conclusive verification of the presence and quantification of stem cells within a sample testis. Biopsies of testes from patients with a one year arrest in spermatogenesis were first minced mechanically into small pieces and then human spermatogonial stem cells (hSSCs) and Sertoli cells isolated by the two- step enzymatic digestion were plated and grown on DSA-Lectin coated dishes in Dulbecco's modified Eagle's medium (DMEM) containing %10 fetal calf serum. Transplantation of human spermatogonial cells into mouse recipient testis was carried out on Day 7 (before colonization) and 2 weeks after culturing (colonization). the effects of different concentrations of spermatogonial cell on quantity of transplantation was assayed during 8 weeks after transplantation. Transplantation of human spermatogonial stem cells to infertile mice model testis indicated that these cells can be observed on the basement membrane of seminiferous tubules in place of spermatogonial stem cells. Proliferation occurs about 4 weeks after transplantation but meiotic differentiation of human spermatogonial cells was not observed in recipient mice testis 8 weeks after transplantation. The difference in donor cell concentration had more effect on colonization of them in mouse recipient testis (p<0.05). Development of germ cell transplantation technique and repopulation of infertile seminiferous tubules is a powerful tool for, study of biology, detecting of activity and quantitative analyses of SSCs. It will be an alternative approach for the preservation of fertility, in the future.