The Effects of Testicular Microenvironment on Proliferation of Spermatogonial Stem Cells in Azoospermia Patients

Objective: This study aims to investigate the testes cultures of patients with previous histories of maturation arrest in spermatogenesis and find the appropriate methods to overcome this problem. Methods: We divided spermatogonial stem cells (SSCs) isolated from testes biopsies into ۳ groups: ۱) culture of SSCs without feeder layer; ۲) co-culture of SSCs with patient-derived Sertoli cells; and ۳) co-culture of SSCs on Sertoli cell feeder layer derived from healthy donors. We calculated the numbers and diameters of stem cell-derived colonies and the percentage of cell viability in the different groups. The presence of SSCs at different culture times was determined by immunochemistry, alkaline phosphatase, and xenotransplantation of SSCs into an azoospermic mouse model. Results: The microenvironment of the feeder layer derived from the patient’s own Sertoli cells produced numerous (۳۶.۱±۴) large colonies (۲۱۳.۲±۱۷ µm) after ۳ weeks of culture. However, the ratio of germ cell-specific expressions of Stra۸ (۲.۳) and Vasa (۲.۲) was more than the pluripotency gene, Nanog (۰.۴۵) in SSCs cultured on the Sertoli cell layer of a healthy person. After xenotransplantation of human SSCs into the testis of an azoospermic mouse model, we observed that the cells grow on basement membrane of seminiferous tubules, which confirmed their nature. Conclusion: SSCs could be co-cultured with Sertoli cells derived from healthy donors in order to overcome the arrest of spermatogenesis observed in the co-culture of SSCs with patient-derived Sertoli cells. The results of the present study indicated that spermatogenesis could possibly be resumed in cancer patients previously treated by chemotherapy and∕or radiotherapy.

کلیدواژه ها:

Sertoli cells ، testis microenvironment ، Spermatogonial stem cells ، ریز محیط بیضه ، سلول های بنیادی اسپرماتوگونی ، سلول های سرتولی