title: Isolation, proliferation, and induction of Bama mini-pig spermatogonial stem cells in vitro.
abstract: Spermatogonial stem cells (SSCs), the unique seed cells of testes, can undergo meiosis and form spermatozoa, thus transmitting genetic information to offspring. Research concerning these cells explores the mechanism underlying spermatogenesis, making possible the induction of their differentiation into spermatozoa in vitro. SSCs have therefore attracted much interest among scientists. Although the proliferation of such cells in vitro has been demonstrated, we are unaware of any long-term laboratory culture of porcine SSCs. The objective of this study was to isolate, characterize, culture, and induce the differentiation of Bama mini-pig SSCs. SSCs were isolated using differential plating and cultured for over 100 days on an STO feeder cell layer without serum. Cell clusters appeared after three passages and continuously formed during subsequent cultivation. Staining showed that these clusters were positive for UCHL1 and CDH1, could be bound by Dolichos biflorus agglutinin, and that some cells expressed OCT4. Ultrastructure observations revealed SSCs in testis tissue to be round in shape, while those cultured in vitro were flat and bound together. Our attempts at inducing differentiation showed that SSCs cultured in vitro could undergo meiosis. In this study, we describe an effective culture system for Bama mini-pig SSCs capable of producing enough cells to establish a platform for further SSC research, such as genetic manipulation or exploration of the mechanism underlying spermatogenesis.
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, China.
authors.forename: H M
authors.initials: HM
authors.lastname: Zhao
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, China.
authors.forename: H
authors.initials: H
authors.lastname: Yang
authors.affiliation: Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, China.
authors.forename: F H
authors.initials: FH
authors.lastname: Luo
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, China.
authors.forename: M X
authors.initials: MX
authors.lastname: Li
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, China.
authors.forename: S
authors.initials: S
authors.lastname: Zhang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, China.
authors.forename: X G
authors.initials: XG
authors.lastname: Yang
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, China.
authors.forename: Y Q
authors.initials: YQ
authors.lastname: Lu
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, China.
authors.forename: S S
authors.initials: SS
authors.lastname: Lu
authors.affiliation: Key Laboratory of National Education Ministry for Mammalian Reproductive Biology and Biotechnology, Inner Mongolia University, Hohhot, China.
authors.forename: Y J
authors.initials: YJ
authors.lastname: Wu
authors.affiliation: State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Laboratory for Animal Reproduction and Biotechnology, Guangxi University, Nanning, China khlu@gxu.edu.cn.
authors.forename: K H
authors.initials: KH
authors.lastname: Lu
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D002449:Cell Aggregation; D002454:Cell Differentiation; D049109:Cell Proliferation; D002469:Cell Separation; D002478:Cells, Cultured; D008297:Male; D008540:Meiosis; D013091:Spermatogenesis; D013093:Spermatogonia / Q000166:cytology*; D013234:Stem Cells / Q000502:physiology*; D013552:Swine; D013556:Swine, Miniature; D013737:Testis / Q000166:cytology
chemical_list: D015415:Biomarkers