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.
% T3 b) C9 D- j  m8 jThis culture system provides a basic platform for understanding the regulation of SSC fate commitment in vitro and for further improving SSC culture medium.
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4 `- v& G( r. C/ O' |# ~' wAddress correspondence and reprint requests to: Takashi Shinohara, Department of Molecular
9 e7 l- m0 l1 i3 y, Y0 YGenetics, Graduate School of Medicine, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto, e% [$ `- M: H4 \6 C2 |/ ^. {
606-8501, Japan3 w) C8 k: g$ R3 G' ^
Tel: 81-75-751-4160; Fax: 81-75-751-4169; E-mail: tshinoha@virus.kyoto-u.ac.jp# L! I+ T- |1 v9 F) O) U
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