Evaluation of Apoptosis-Related Genes and Hormone Secretion Profiles Using Three Dimensional Culture System of Human Testicular Organoids

Background: In reproductive biology, testicular organoids can be used to treat infertility and study development spermatogonial stem cells (SSCs) differentiation. Generating organoid from primary is challenging. this study, were created using human evaluated the apoptotic gene expression hormone secretion profiles of organoids. Materials Methods: Primary isolated 2-step enzymatic digestion three brain-dead donors. Immunocytochemistry flow cytometry analyses performed confirm SSCs. Isolated cultured in experimental groups: control group (2 dimensional (2D)), 1 (organoid culture after 2D culture), 2 immediately digestion). Testicular DMEM/F-12 media supplemented with follicle-stimulating (FSH) fetal bovine serum (FBS) for four weeks. After 24 hours weeks culture, reverse transcription quantitative real-time PCR (RT-qPCR) was investigate relative genes (caspase 3, 9, Bax, Bcl-2). At hours, two weeks, enzyme-linked immunoassay (ELISA) determine testosterone inhibin B concentrations. Light microscopy toluidine blue staining also morphological analysis. Results: RT-qPCR results revealed that pro-apoptotic Bax) levels highest h culture. contrast, level Bcl-2 (anti-apoptotic) lower compared other groups. The decreased a time-dependent manner during cultivation. According evaluations, are compact, spherical structures elongated organized along their border. Conclusion: Our findings system maintained hormonal secretory abilities, demonstrating function Sertoli Leydig absence testis-specific environments.