title: A novel feeder-free culture system for expansion of mouse spermatogonial stem cells.
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.
authors.affiliation: Department of Stem Cell Biology, School of Medicine, Konkuk University, Seoul 143-701, Korea ; Center for Stem Cell Research, Institute of Advanced Biomedical Science, Konkuk University, Seoul 143-701, Korea.
authors.forename: Na Young
authors.initials: NY
authors.lastname: Choi
authors.forename: Yo Seph
authors.initials: YS
authors.lastname: Park
authors.forename: Jae-Sung
authors.initials: JS
authors.lastname: Ryu
authors.forename: Hye Jeong
authors.initials: HJ
authors.lastname: Lee
authors.forename: Marcos J
authors.initials: MJ
authors.lastname: Araúzo-Bravo
authors.forename: Kisung
authors.initials: K
authors.lastname: Ko
authors.forename: Dong Wook
authors.initials: DW
authors.lastname: Han
authors.forename: Hans R
authors.initials: HR
authors.lastname: Schöler
authors.forename: Kinarm
authors.initials: K
authors.lastname: Ko
mesh_terms: D053687:Adult Stem Cells / Q000166:cytology* / Q000502:physiology* / Q000637:transplantation; D000818:Animals; D049109:Cell Proliferation; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry; D003584:Cytological Techniques / Q000191:economics / Q000379:methods*; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D013093:Spermatogonia / Q000166:cytology*
chemical_list: D003470:Culture Media