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

M Kanatsu-Shinohara, K Inoue, N Ogonuki… - Biology of …, 2011 - academic.oup.com
M Kanatsu-Shinohara, K Inoue, N Ogonuki, H Morimoto, A Ogura, T Shinohara
Biology of reproduction, 2011academic.oup.com
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 for long-term
propagation of SSCs. In addition to the SSC self-renewal factors, including glial cell line-
derived neurotrophic factor, supplementation with fetuin and lipid-associated molecules was …
Abstract
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 for long-term propagation of SSCs. In addition to the SSC self-renewal factors, including glial cell line-derived neurotrophic factor, supplementation with fetuin and lipid-associated molecules was required to drive SSC proliferation in vitro. Cultured cells proliferated for at least 6 mo at a rate comparable to that of serum-supplemented cultured cells. However, germline potential was reduced under serum- and feeder-free conditions, as indicated by 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. This culture system provides a basic platform for understanding the regulation of SSC fate commitment in vitro and for improving SSC culture medium.
Oxford University Press