2014年10月1日
Improved serum- and feeder-free culture of mouse germline stem cells
Biology of Reproduction
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- 巻
- 91
- 号
- 4
- 開始ページ
- 88
- 終了ページ
- 記述言語
- 英語
- 掲載種別
- 研究論文(学術雑誌)
- DOI
- 10.1095/biolreprod.114.122317
- 出版者・発行元
- Society for the Study of Reproduction
Spermatogonial stem cells (SSCs) undergo self-renewal division, which can be recapitulated in vitro. Attempts to establish serum-free culture conditions for SSCs have met with limited success. Although we previously reported that SSCs can be cultured without serum on laminin-coated plates, the growth rate and SSC concentration were relatively low, which made it inefficient for culturing large numbers of SSCs. In this study, we report on a novel culture medium that showed improved SSC maintenance. We used Iscove modified Dulbecco medium, supplemented with lipid mixture, fetuin, and knockout serum replacement. In the presence of glial cell line-derived neurotrophic factor (GDNF) and fibroblast growth factor 2 (FGF2), SSCs cultured on laminin-coated plates could proliferate for more than 5 mo and maintained normal karyotype and androgenetic DNA methylation patterns in imprinted genes. Germ cell transplantation showed that SSCs in the serum-free medium proliferated more actively than those in the serumsupplemented medium and that the frequency of SSCs was comparable between the two culture media. Cultured cells underwent germline transmission. Development of a new serumand feeder-free culture method for SSCs will facilitate studies into the effects of microenvironments on self-renewal and will stimulate further improvements to derive SSC cultures from different animal species.
- リンク情報
- ID情報
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- DOI : 10.1095/biolreprod.114.122317
- ISSN : 1529-7268
- ISSN : 0006-3363
- PubMed ID : 25210127
- SCOPUS ID : 84930515659