a comparative analysis of culture systems with human amniotic mesenchymal stem cells

Background: a indirect co-culture using mesh system is commonly employed to maintain spermatogenesis in cancer patients undergoing chemotherapy and radiation. this study aimed to investigate the co-culturing of mouse spermatogonial stem cells (sscs) with human amniotic mesenchymal stem/stromal cells (hamscs) in an optimized environment. methods: sscs from 3-6-day-old mice (n=10) were indirectly co-cultured with hamscs via mesh for two weeks. three groups evaluated: control, sscs with conditioned media, and sscs indirectly co-cultured with hamscs. gene expression analyzed for plzf, c-kit, sycp3, and prm1. immunohistochemistry assessed plzf, and flow cytometry evaluated c-kit and plzf. results: showed a twofold increase in plzf-positive cells after 14 days of culture (76.47%, p≤0.05), with a significant elevation in plzf gene expression observed in the conditioned media group (188.1 ± 65%, p≤0.05). conversely, the expression of the c-kit gene decreased significantly in both the conditioned media and indirect co-culture using mesh groups. notably, sycp3 and prm1 expression levels significantly increased in the conditioned media group compared to the control. these findings suggest the potential of conditioned media as a novel feeder for promoting in vitro mouse spermatogenesis. conclusion: our results demonstrate that the inclusion of growth factors, such as gdnf and bmp-4, along with conditioned media and an indirect co-culture using mesh system utilizing meshes with sscs, significantly enhances ssc proliferation and differentiation. the optimized conditions media provided by hamscs offer a superior feeder compared to traditional indirect co-culture using mesh systems for promoting both the proliferation and differentiation of sscs.