Establishment of alternative culture method for spermatogonial stem cells using knockout serum replacement.

Since spermatogonial stem cells (SSCs) are capable of both self-renewal and differentiation to daughter cells for subsequent spermatogenesis, the development of an efficient in vitro culture system is essential for studies related to spermatogenesis. Although the currently available system is serum-...

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Autores principales:	Keisuke Aoshima, Ai Baba, Yoshinori Makino, Yuki Okada
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2013
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7c664400704741688370d2b3e25a2fbe
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spelling	oai:doaj.org-article:7c664400704741688370d2b3e25a2fbe2021-11-18T08:49:16ZEstablishment of alternative culture method for spermatogonial stem cells using knockout serum replacement.1932-620310.1371/journal.pone.0077715https://doaj.org/article/7c664400704741688370d2b3e25a2fbe2013-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/24204931/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203Since spermatogonial stem cells (SSCs) are capable of both self-renewal and differentiation to daughter cells for subsequent spermatogenesis, the development of an efficient in vitro culture system is essential for studies related to spermatogenesis. Although the currently available system is serum-free and contains only chemically-defined components, it highly relies upon bovine serum albumin (BSA), a component with batch-to-batch quality variations similar to those of fetal bovine serum. Thus, we searched for an alternative BSA-free culture system that preserved the properties of SSCs. In this study, we utilized Knockout Serum Replacement (KSR) in the SSC culture medium, as a substitute for BSA. The results demonstrated that KSR supported the continuous growth of SSCs in vitro and the SSC activity in vivo without BSA, in a feeder-cell combination with mouse embryonic fibroblasts. The addition of BSA to KSR further facilitated cell cycle progression, whereas a transplantation assay revealed that the addition of BSA did not affect the number of SSCs in vivo. The combination of KSR with BSA also allowed the elimination of GFRA1 and FGF2, and the reduction of the GDNF concentration from 20 ng/ml to 5 ng/ml, while maintaining the growth rate and the expression of SSC markers. Furthermore, KSR was also useful with SSCs from non-DBA/2 strains, such as C57BL/6 and ICR. These results suggested that KSR is an effective substitute for BSA for long-term in vitro cultures of SSCs. Therefore, this method is practical for various studies related to SSCs, including spermatogenesis and germ stem cell biology.Keisuke AoshimaAi BabaYoshinori MakinoYuki OkadaPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 8, Iss 10, p e77715 (2013)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Keisuke Aoshima Ai Baba Yoshinori Makino Yuki Okada Establishment of alternative culture method for spermatogonial stem cells using knockout serum replacement.
description	Since spermatogonial stem cells (SSCs) are capable of both self-renewal and differentiation to daughter cells for subsequent spermatogenesis, the development of an efficient in vitro culture system is essential for studies related to spermatogenesis. Although the currently available system is serum-free and contains only chemically-defined components, it highly relies upon bovine serum albumin (BSA), a component with batch-to-batch quality variations similar to those of fetal bovine serum. Thus, we searched for an alternative BSA-free culture system that preserved the properties of SSCs. In this study, we utilized Knockout Serum Replacement (KSR) in the SSC culture medium, as a substitute for BSA. The results demonstrated that KSR supported the continuous growth of SSCs in vitro and the SSC activity in vivo without BSA, in a feeder-cell combination with mouse embryonic fibroblasts. The addition of BSA to KSR further facilitated cell cycle progression, whereas a transplantation assay revealed that the addition of BSA did not affect the number of SSCs in vivo. The combination of KSR with BSA also allowed the elimination of GFRA1 and FGF2, and the reduction of the GDNF concentration from 20 ng/ml to 5 ng/ml, while maintaining the growth rate and the expression of SSC markers. Furthermore, KSR was also useful with SSCs from non-DBA/2 strains, such as C57BL/6 and ICR. These results suggested that KSR is an effective substitute for BSA for long-term in vitro cultures of SSCs. Therefore, this method is practical for various studies related to SSCs, including spermatogenesis and germ stem cell biology.
format	article
author	Keisuke Aoshima Ai Baba Yoshinori Makino Yuki Okada
author_facet	Keisuke Aoshima Ai Baba Yoshinori Makino Yuki Okada
author_sort	Keisuke Aoshima
title	Establishment of alternative culture method for spermatogonial stem cells using knockout serum replacement.
title_short	Establishment of alternative culture method for spermatogonial stem cells using knockout serum replacement.
title_full	Establishment of alternative culture method for spermatogonial stem cells using knockout serum replacement.
title_fullStr	Establishment of alternative culture method for spermatogonial stem cells using knockout serum replacement.
title_full_unstemmed	Establishment of alternative culture method for spermatogonial stem cells using knockout serum replacement.
title_sort	establishment of alternative culture method for spermatogonial stem cells using knockout serum replacement.
publisher	Public Library of Science (PLoS)
publishDate	2013
url	https://doaj.org/article/7c664400704741688370d2b3e25a2fbe
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Establishment of alternative culture method for spermatogonial stem cells using knockout serum replacement.

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