title: Isolation and Identification of Prepubertal Buffalo (Bubalus bubalis) Spermatogonial Stem Cells.
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.
authors.affiliation: State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530005, China.
authors.forename: Wanyou
authors.initials: W
authors.lastname: Feng
authors.affiliation: State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530005, China.|Center of Reproduction of Nanxishan Hospital, Guilin 541002, China.
authors.forename: Shibei
authors.initials: S
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530005, China.|Bacgiang Agriculture and Forestry University, Bacgiang 220000 Vietnam.
authors.forename: Dagiang
authors.initials: D
authors.lastname: Do
authors.affiliation: State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530005, China.
authors.forename: Qinyou
authors.initials: Q
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530005, China.
authors.forename: Yanfei
authors.initials: Y
authors.lastname: Deng
authors.affiliation: State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530005, China.
authors.forename: Xiaocan
authors.initials: X
authors.lastname: Lei
authors.affiliation: State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530005, China.
authors.forename: Chan
authors.initials: C
authors.lastname: Luo
authors.affiliation: State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530005, China.
authors.forename: Ben
authors.initials: B
authors.lastname: Huang
authors.affiliation: State Key Laboratory of Conservation and Untilization of Subtropical Agro-Bioresources, Guangxi University, Nanning 530005, China.
authors.forename: Deshun
authors.initials: D
authors.lastname: Shi
keywords: Buffalo; Germ Cell Marker; Proliferation; Spermatogonial Stem Cell