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.
! C0 f/ O4 T( }- y+ sThis culture system provides a basic platform for understanding the regulation of SSC fate commitment in vitro and for further improving SSC culture medium.( @2 A/ a! U# D: N4 t

- B* Z- |# D$ B2 U/ N! P; m3 GAddress correspondence and reprint requests to: Takashi Shinohara, Department of Molecular% k* I& n1 g- z8 L1 `: H$ ~/ M+ `" u
Genetics, Graduate School of Medicine, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto; E- b/ l/ A9 X# \1 g
606-8501, Japan
! t0 {5 k: N, }5 u5 o/ Z6 kTel: 81-75-751-4160; Fax: 81-75-751-4169; E-mail: tshinoha@virus.kyoto-u.ac.jp' h) l  B. a: `. I

) ]9 p+ Z1 F7 ]5 d! vFull paper