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.
* \( ]/ e: s1 y" ?This culture system provides a basic platform for understanding the regulation of SSC fate commitment in vitro and for further improving SSC culture medium.
, [  }5 c; y5 ?: h( s, f8 U
+ ]! N- q5 d% `! \Address correspondence and reprint requests to: Takashi Shinohara, Department of Molecular2 X. r; i1 v3 l$ c* m4 @9 A
Genetics, Graduate School of Medicine, Kyoto University, Yoshida Konoe, Sakyo-ku, Kyoto* a: K# V7 p+ C  f
606-8501, Japan+ i( T1 k) j% A" z
Tel: 81-75-751-4160; Fax: 81-75-751-4169; E-mail: tshinoha@virus.kyoto-u.ac.jp
( y. q( ^! p/ H, v" e: G; G
' T1 ?7 k# o" Z3 R3 h/ iFull paper