J Vet Sci.  2018 Sep;19(5):592-599. 10.4142/jvs.2018.19.5.592.

In vitro differentiation of spermatogonial stem cells using testicular cells from Guangxi Bama mini-pig

• Zhao H
• Nie J
• Zhu X
• Lu Y
• Liang X
• Xu H
• Yang X
• Zhang Y
• Lu K
• Lu S

• KMID: 2420927
• DOI: http://doi.org/10.4142/jvs.2018.19.5.592

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.

Keyword

mini pig; spermatogenesis; stem cell; testicular

MeSH Terms

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
Acrosin
Activins
Alkaline Phosphatase
Bone Morphogenetic Protein 4
Follicle Stimulating Hormone
Testosterone
Tretinoin