Molecules and Cells

Korean Society for Molecular and Cellular Biology (한국분자세포생물학회)
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A Novel Feeder-Free Culture System for Expansion of Mouse Spermatogonial Stem Cells

  • Choi, Na Young (Department of Stem Cell Biology, School of Medicine, Konkuk University) ;
  • Park, Yo Seph (Department of Stem Cell Biology, School of Medicine, Konkuk University) ;
  • Ryu, Jae-Sung (Center for Stem Cell Research, Institute of Advanced Biomedical Science, Konkuk University) ;
  • Lee, Hye Jeong (Department of Stem Cell Biology, School of Medicine, Konkuk University) ;
  • Arauzo-Bravo, Marcos J. (Group of Computational Biology and Bioinformatics, Biodonostia Health Research Institute) ;
  • Ko, Kisung (Department of Medicine, Medical Research Institute, College of Medicine, Chung-Ang University) ;
  • Han, Dong Wook (Department of Stem Cell Biology, School of Medicine, Konkuk University) ;
  • Scholer, Hans R. (Department of Cell and Developmental Biology, Max Planck Institute for Molecular Biomedicine) ;
  • Ko, Kinarm (Department of Stem Cell Biology, School of Medicine, Konkuk University)
  • Received : 2014.04.04
  • Accepted : 2014.04.28
  • Published : 2014.06.30
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Abstract

Spermatogonial stem cells (SSCs, also called germline stem cells) are self-renewing unipotent stem cells that produce differentiating germ cells in the testis. SSCs can be isolated from the testis and cultured in vitro for long-term periods in the presence of feeder cells (often mouse embryonic fibroblasts). However, the maintenance of SSC feeder culture systems is tedious because preparation of feeder cells is needed at each subculture. In this study, we developed a Matrigel-based feeder-free culture system for long-term propagation of SSCs. Although several in vitro SSC culture systems without feeder cells have been previously described, our Matrigel-based feeder-free culture system is time- and cost-effective, and preserves self-renewability of SSCs. In addition, the growth rate of SSCs cultured using our newly developed system is equivalent to that in feeder cultures. We confirmed that the feeder-free cultured SSCs expressed germ cell markers both at the mRNA and protein levels. Furthermore, the functionality of feeder-free cultured SSCs was confirmed by their transplantation into germ cell-depleted mice. These results suggest that our newly developed feeder-free culture system provides a simple approach to maintaining SSCs in vitro and studying the basic biology of SSCs, including determination of their fate.

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References

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