In vitro differentiation of spermatogonial stem cells using testicular cells from Guangxi Bama mini-pig
10.4142/jvs.2018.19.5.592
- Author:
Huimin ZHAO
1
;
Junyu NIE
;
Xiangxing ZHU
;
Yangqing LU
;
Xingwei LIANG
;
Huiyan XU
;
Xiaogan YANG
;
Yunkai ZHANG
;
Kehuan LU
;
Shengsheng LU
Author Information
1. State 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, Guangxi University, Nanning 530005, China. shengshenglu@sina.com
- Publication Type:Original Article
- Keywords:
mini pig;
spermatogenesis;
stem cell;
testicular
- MeSH:
Acrosin;
Activins;
Alkaline Phosphatase;
Bone Morphogenetic Protein 4;
Dolichos;
Follicle Stimulating Hormone;
Humans;
In Vitro Techniques;
Infant;
Infant, Newborn;
Spermatogenesis;
Stem Cells;
Swine;
Testis;
Testosterone;
Tretinoin;
Up-Regulation
- From:Journal of Veterinary Science
2018;19(5):592-599
- CountryRepublic of Korea
- Language:English
-
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-dimensional 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.
