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Title: Long-term culture of undifferentiated spermatogonia isolated from immature and adult bovine testes
Authors: Suyatno
Kitamura, Yuka
Ikeda, Shuntaro  kyouindb  KAKEN_id
Minami, Naojiro  kyouindb  KAKEN_id
Yamada, Masayasu  kyouindb  KAKEN_id
Imai, Hiroshi
Author's alias: 南, 直治郎
山田, 雅保
今井, 裕
Keywords: adult male germ cells
pluripotent stem cells
spermatogonial stem cells
Issue Date: Mar-2018
Publisher: Wiley-Blackwell
Journal title: Molecular Reproduction and Development
Volume: 85
Issue: 3
Start page: 236
End page: 249
Abstract: Undifferentiated spermatogonia eventually differentiate in the testis to produce haploid sperm. Within this cell population, there is a small number of spermatogonial stem cells (SSCs). SSCs are rare cells in the testis, and their cellular characteristics are poorly understood. Establishment of undifferentiated cell line would provide an indispensable tool for studying their biological nature and spermiogenesis/spermatogenesis in vitro. However, there have been few reports on the long-term culture of undifferentiated spermatogonia in species other than rodents. Here, we report the derivation and long-term in vitro culture of undifferentiated spermatogonia cell lines from immature and adult bovine testes. Cell lines from immature testes were maintained in serum-free culture conditions in the presence of glial-cell-line-derived neurotropic factor (GDNF) and bovine leukemia inhibitory factor (bLIF). These cell lines have embryonic stem (ES)-like cell morphology, express pluripotent-stem-cell-specific and germ-cell-specific markers at the protein and mRNA levels, and contributed to the inner cell mass (ICM) of embryos in the blastocyst stage. Meanwhile, cell lines established from adult testes were maintained in low-serum media in the presence of 6-bromoindirubin-3′-oxime (BIO). These cell lines have characteristics resembling those of previously reported male mouse germ cell lines as confirmed by their botryoidally aggregated morphology, as well as the expression of germ-cell-specific markers and pluripotent stem cell markers. These findings could be useful for the development of long-term culture of undifferentiated spermatogonia, which could aid in conservation of species and improvement of livestock production through genome editing technology.
Description: ウシ精巣に由来する多能性生殖幹細胞株の樹立 --精子になるはずのものが多能性細胞に--. 京都大学プレスリリース. 2018-03-08.
Rights: This is the accepted version of the following article: [Suyatno, Kitamura Y, Ikeda S, Minami N, Yamada M, Imai H. Long-term culture of undifferentiated spermatogonia isolated from immature and adult bovine testes. Mol Reprod Dev. 2018;85:236–249], which has been published in final form at This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Self-Archiving.
The full-text file will be made open to the public on 13 MAR 2019 in accordance with publisher's 'Terms and Conditions for Self-Archiving'.
This is not the published version. Please cite only the published version. この論文は出版社版でありません。引用の際には出版社版をご確認ご利用ください。
DOI(Published Version): 10.1002/mrd.22958
PubMed ID: 29480937
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