Serum- and feeder-free culture of mouse germline stem cells1

chiangmax

Spermatogonial stem cells (SSCs) undergo self-renewal divisions to support spermatogenesis. Although several in vitro SSC culture systems have been developed, these systems include serum or fibroblast feeders, which complicate SSC self-renewal analyses. Here we developed a serum and feeder-free culture system, in which SSCs propagated for long-term. In addition to the SSC self-renewal factors including glial cell line-derived neurotrophic factor (GDNF), supplementation with fetuin and lipid-associated molecules was required to drive SSC proliferation in vitro. Cultured cells proliferated for at least 6 months at a rate comparable to that of serum-supplemented cultured cells. However, germline potential was reduced under serumand feeder-free conditions, because we observed a lower SSC frequency after germ cell transplantation. Nevertheless, the cultured cells completed spermatogenesis and produced offspring following spermatogonial transplantation into seminiferous tubules of infertile mice.
1 z/ _7 ]; P% k7 z& tThis culture system provides a basic platform for understanding the regulation of SSC fate commitment in vitro and for further improving SSC culture medium.8 K  X3 H* Y7 [2 a* A
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Address correspondence and reprint requests to: Takashi Shinohara, Department of Molecular0 a: P1 U% T/ G: |4 E1 s3 V
Genetics, Graduate School of Medicine, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto. T7 y4 j7 X( s) l
606-8501, Japan
0 E) G* B/ _+ s" M5 m" y- bTel: 81-75-751-4160; Fax: 81-75-751-4169; E-mail: tshinoha@virus.kyoto-u.ac.jp
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