<> "The repository administrator has not yet configured an RDF license."^^ . <> . . "Identification and Pluripotency of Mouse\r\nSpermatogonial Stem Cells"^^ . "Spermatogonial stem cells (SSCs) are located on the basal membrane in seminiferous tubules of the testis and are responsible for sperm production during the male’s life. Although the morphology and kinetics of spermatogonia has been extensively characterized, there is still a lack of specific cellular, morphological and biochemical criteria for SSCs. However, Fluorescence-activated cell sorting (FACS), magnetic-activated cell sorting (MACS), matrix selection and morphology-based selection are some of the useful methods for the isolation of SSCs. \r\nIn the current study, we demonstrated that using cell sorting approaches are dispensable for the generation of mouse SSCs cultures. We identified a simple and highly reproducible protocol for the isolation of SSCs with morphology-based selection and successfully established two types of SSCs (Type I and Type II). Using co-culture systems with different mitomycin-C treated feeder layers in long-term culture, we established the optimal conditions for the cultivation and gene expansion of these two types of SSCs. We successfully expanded Type I SSCs on mouse embryonic fibroblasts (MEFs) and Type II SSCs on both SNL and primary testicular stroma cells (TSCs) feeders. Electron microscopic analysis revealed that Type I SSCs display a similar morphology with SSCs in-vivo high nucleus/cytoplasm ratio, while Type II SSCs have a different morphology small nucleus/cytoplasm ratio. Immunocytochemistry, FACS and Fluidigm real-time RT-PCR results showed that Type I SSCs clearly express germ cells markers while Type II SSCs only partially express the typical germ cell profile of SSCs. Following the transplantation of Type II SSCs to busulphan-treated NOD/ SCID mice, we observed a localization of GFP-labeled cells in the basal compartment of the seminiferous tubule. Furthermore, GFP labeled sperms were detected in epididymis. Among the most obvious molecular differences between Type I and Type II SSCs were their reprogramming capacity to mouse embryonic stem cells-(ES-) like cells that occurred only on the critical time window after initiation of Type I SSCs culture. Although testis-derived ES-like cells have been obtained in several studies, the time window for the shift to pluripotency was not clearly determined. In our experiments, we observed that the spontaneous appearance of germline-derived ES-like cells from both neonate and adult Type I cells occurred only during a special time window (41 until 125 days) after initiation of Type I cells from neonate and adult promoter-reporter Oct4-GFP transgenic mice. The generated ES-like cells expressed pluripotency markers, differentiated into all three germ lineages, formed complex teratoma after transplantation in SCID mice and produced chimeric mice. Although the exact mechanism of the development of ES-like cells from GSCs is still unclear, this new information could provide an ideal strategy for scheduling natural conversion mechanisms of ES-like cells from mouse testes. Therefore, the two different types of SSCs could provide an ideal cell system for studying both pluripotency and in-vitro differentiation of SSCs to sperm and also provide a new strategy for isolation of SSCs from neonatal and adult mice by morphology-based selection."^^ . "2014" . . . . . . . "Hossein"^^ . "Azizi"^^ . "Hossein Azizi"^^ . . . . . . "Identification and Pluripotency of Mouse\r\nSpermatogonial Stem Cells (PDF)"^^ . . . "Hossein Azizi.pdf"^^ . . . "Identification and Pluripotency of Mouse\r\nSpermatogonial Stem Cells (Other)"^^ . . . . . . "lightbox.jpg"^^ . . . "Identification and Pluripotency of Mouse\r\nSpermatogonial Stem Cells (Other)"^^ . . . . . . "preview.jpg"^^ . . . "Identification and Pluripotency of Mouse\r\nSpermatogonial Stem Cells (Other)"^^ . . . . . . "medium.jpg"^^ . . . "Identification and Pluripotency of Mouse\r\nSpermatogonial Stem Cells (Other)"^^ . . . . . . "small.jpg"^^ . . . "Identification and Pluripotency of Mouse\r\nSpermatogonial Stem Cells (Other)"^^ . . . . . . "indexcodes.txt"^^ . . "HTML Summary of #17280 \n\nIdentification and Pluripotency of Mouse \nSpermatogonial Stem Cells\n\n" . "text/html" . . . "500 Naturwissenschaften und Mathematik"@de . "500 Natural sciences and mathematics"@en . .