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      Genetic and epigenetic properties of mouse male germline stem cells during long-term culture.

      Development (Cambridge, England)

      Animals, Cell Culture Techniques, Cell Proliferation, Cell Survival, physiology, Chromosomes, Mammalian, genetics, Cytogenetic Analysis, Electrophoresis, Gel, Pulsed-Field, Embryo Transfer, Epigenesis, Genetic, Fluorescence, Glial Cell Line-Derived Neurotrophic Factor, Male, Mice, Mice, Transgenic, Restriction Mapping, Reverse Transcriptase Polymerase Chain Reaction, Spermatogenesis, Spermatogonia, cytology, Stem Cells, Telomerase, metabolism, Telomere, Testis

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          Although stem cells are believed to divide infinitely by self-renewal division, there is little evidence that demonstrates their infinite replicative potential. Spermatogonial stem cells are the founder cell population for spermatogenesis. Recently, in vitro culture of spermatogonial stem cells was described. Spermatogonial stem cells can be expanded in vitro in the presence of glial cell line-derived neurotrophic factor (GDNF), maintaining the capacity to produce spermatogenesis after transplantation into testis. Here, we examined the stability and proliferative capacity of spermatogonial stem cells using cultured cells. Spermatogonial stem cells were cultured over 2 years and achieved approximately 10(85)-fold expansion. Unlike other germline cells that often acquire genetic and epigenetic changes in vitro, spermatogonial stem cells retained the euploid karyotype and androgenetic imprint during the 2-year experimental period, and produced normal spermatogenesis and fertile offspring. However, the telomeres in spermatogonial stem cells gradually shortened during culture, suggesting that they are not immortal. Nevertheless, the remarkable stability and proliferative potential of spermatogonial stem cells suggest that they have a unique machinery to prevent transmission of genetic and epigenetic damages to the offspring, and these characteristics make them an attractive target for germline modification.

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