Therapeutic efficacy of mesenchymal stem cells for the restoration of dysfunctional rat testes

Introduction Chemotherapy, especially with alkylating agents, can impair spermatogonial stem cells in young males, leading to infertility. This study evaluates the therapeutic efficacy of bone-marrow-derived Mesenchymal Stem Cells (BM-MSCs) in restoring testicular function of the dysfunctional testes in a chemo-ablated infertile rat model. Method Wistar male rats were randomly divided into three groups: the first control group without any treatment; the second group was infertile chemo-ablated, generated through the intraperitoneal injections of 15 mg/kg of Busulfan, twice with a two-week interval; and the third group was chemo-ablated and subsequently treated with intra-testicular injections of enriched MSCs. The BM-MSCs were cultured and sorted using magnetic-activated cell sorting (MACS), and purity was confirmed by Fluorescence-Activated Cell Sorting (FACS) analysis of positive and negative cell-surface markers. Purified MSCs were injected into the testes of the infertile rats. Results were evaluated after 2-10 weeks of treatment. After 2 weeks of MSC treatment, mating trials for these males were conducted every 2 weeks, and their testes, as well as serum samples; were collected for therapeutic assessment of MSCs. Results Histological assessments of the excised testes revealed a gradual, significant improvement within 10 weeks of treatment. Additionally, the expression levels of stem cell and sperm maturation markers were elevated in the testes of treated animals, indicating recovery of spermatogenesis. Hormonal analyses demonstrated normalisation of FSH and Testosterone levels after treatment. Mating studies also demonstrated that the MSC-treated group delivered 7-8 pups per litter, whereas chemo-ablated animals remained totally infertile. Conclusions Overall findings support MSC transplantation as a promising approach to restore testicular function and fertility, inspiring continued exploration in regenerative medicine.