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Many of the testicular cancer-survived patients, treated with chemotherapeutic drugs, show infertility, pre and postimplantation loss, and germ cell abnormality. Studies examining the negative effects of chemotherapeutic drugs on testicular germ cells are ongoing; however, information on the stemness properties and proteomic profiles of these cells are lacking. This study investigated the effects of chemotherapeutic drugs etoposide, cisplatin, bleomycin, and their combination (BEP) on the physiology and stem cell activity of mouse germ cells in vitro. Our results showed that treatment with the abovementioned drugs affected germ cell viability and decreased the number of proliferating germ cells significantly at specific concentrations (0.05 µM etoposide, 1 µM cisplatin, 10 µM bleomycin, and 0.1 µM BEP), which maintained a survival rate of &gt;90%. We also observed a significantly higher percentage of apoptotic cells and alterations in the expression of undifferentiated and differentiated spermatogonia-related genes and marker proteins in germ cells exposed to abovementioned concentrations of the drugs. Next, we performed germ cell transplantation into recipient mice and observed a remarkable reduction in stemness properties of spermatogonial stem cells at these concentrations. Based on these results, we assessed the levels of differentially expressed proteins by performing proteomic analysis. We found that treatment with the abovementioned drugs induced cell damage, oxidative stress, metabolic disruption, and immune deficiency which may promote tumor regeneration, cytotoxicity, infertility, and transgenerational cellular function transmission. Thus, this study provides information about the chemotherapy-induced recurrent destruction and thereby can lead possible changes in medication.

Chemotherapeutic Drugs Alter Functional Properties and Proteome of Mouse Testicular Germ Cells In Vitro