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J Vet Sci 2018; 19(5): 592-599  https://doi.org/10.4142/jvs.2018.19.5.592
In vitro differentiation of spermatogonial stem cells using testicular cells from Guangxi Bama mini-pig
Huimin Zhao1,2, Junyu Nie1, Xiangxing Zhu1, Yangqing Lu1, Xingwei Liang1, Huiyan Xu1, Xiaogan Yang1, Yunkai Zhang2, Kehuan Lu1, Shengsheng Lu1,*
1State Key Laboratory for Conservation and Utilization of Subtropical Agro-bioresources, Guangxi High Education Key Laboratory for Animal Reproduction and Biotechnology, College of Animal Science and Technology and 2College of Life Science and Technology, Guangxi University, Nanning 530005, China
Correspondence to: Tel: +86-771-3235707; Fax: +86-771-3238064; E-mail: shengshenglu@sina.com
Received: August 1, 2017; Revised: April 28, 2018; Accepted: May 9, 2018; Published online: September 30, 2018.
Abstract
In this study, we attempted to establish a culture system for in vitro spermatogenesis from spermatogonial stem cells (SSCs) of Bama mini-pig. Dissociated testicular cells from 1-month-old pigs were co-cultured to mimic in vivo spermatogenesis. The testicular cells were seeded in minimum essential medium alpha (α-MEM) supplemented with Knockout serum replacement (KSR). Three-imensional colonies formed after 10 days of culture. The colonies showed positive staining for SSC-associated markers such as UCHL1, PLZF, THY1, OCT4, Dolichos biflorus agglutinin, and alkaline phosphatase. Induction of SSCs was performed in α-MEM + KSR supplemented with retinoic acid, bone morphogenetic protein 4, activin A, follicle-stimulating hormone, or testosterone. The results showed that STRA8, DMC1, PRM1, and TNP1 were upregulated significantly in the colonies after induction compared to that in testis from 1-month-old pigs, while expression levels of those genes were significantly low compared to those in 2-month-old testis. However, upregulation of ACROSIN was not significant. Replacement of α-MEM and KSR with Iscove’s modified Dulbecco’s medium and fetal bovine serum did not upregulate expression of these genes significantly. These results indicate that SSCs of Bama mini-pig could undergo differentiation and develop to a post-meiotic stage in α-MEM supplemented with KSR and induction factors.
Keywords: mini pig, spermatogenesis, stem cell, testicular


© 2018 The Korean Society of Veterinary Science.

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