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http://dx.doi.org/10.5713/ajas.15.0592

Isolation and Identification of Prepubertal Buffalo (Bubalus bubalis) Spermatogonial Stem Cells  

Feng, Wanyou (State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University)
Chen, Shibei (State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University)
Do, Dagiang (State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University)
Liu, Qinyou (State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University)
Deng, Yanfei (State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University)
Lei, Xiaocan (State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University)
Luo, Chan (State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University)
Huang, Ben (State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University)
Shi, Deshun (State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University)
Publication Information
Asian-Australasian Journal of Animal Sciences / v.29, no.10, 2016 , pp. 1407-1415 More about this Journal
Abstract
Isolation and culture of spermatogonial stem cells (SSCs) are attractive for production of genetic modified offspring. In the present study, buffalo spermatogonial stem-like cells were isolated, cultured and expression pattern of different germ cell marker genes were determined. To recover spermatogonia, testes from age 3 to 7 months of buffalo were decapsulated, and seminiferous tubules were enzymatically dissociated. Two types of cells, immature sertoli cell and type A spermatogonia were observed in buffalo testes in this stage. Germ cell marker genes, OCT3/4 (Pou5f1), THY-1, c-kit, PGP9.5 (UCHL-1) and Dolichos biflorus agglutinin, were determined to be expressed both in mRNA and protein level by reverse transcription polymerase chain reaction and immunostaining in buffalo testes and buffalo spermatogonial stem-like cells, respectively. In the following, when the isolated buffalo buffalo spermatogonial stem-like cells were cultured in the medium supplemented 2.5% fetal bovine serum and 40 ng/mL glial cell-derived neurotrophic factor medium, SSCs proliferation efficiency and colony number were significantly improved than those of other groups (p<0.05). These findings may help in isolation and establishing long term in vitro culture system for buffalo spermatogonial stem-like cells, and accelerating the generation of genetic modified buffaloes.
Keywords
Buffalo; Spermatogonial Stem Cell; Germ Cell Marker; Proliferation;
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