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

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

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 spermatog...

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Main Authors: Feng, Wanyou, Chen, Shibei, Do, Dagiang, Liu, Qinyou, Deng, Yanfei, Lei, Xiaocan, Luo, Chan, Huang, Ben, Shi, Deshun
Format: Online
Language:English
Published: Asian-Australasian Association of Animal Production Societies (AAAP) and Korean Society of Animal Science and Technology (KSAST) 2016
Online Access:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5003965/
id pubmed-5003965
recordtype oai_dc
spelling pubmed-50039652016-10-01 Isolation and Identification of Prepubertal Buffalo (Bubalus bubalis) Spermatogonial Stem Cells Feng, Wanyou Chen, Shibei Do, Dagiang Liu, Qinyou Deng, Yanfei Lei, Xiaocan Luo, Chan Huang, Ben Shi, Deshun Article 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. Asian-Australasian Association of Animal Production Societies (AAAP) and Korean Society of Animal Science and Technology (KSAST) 2016-10 2015-11-16 /pmc/articles/PMC5003965/ /pubmed/26954139 http://dx.doi.org/10.5713/ajas.15.0592 Text en Copyright © 2016 by Asian-Australasian Journal of Animal Sciences This is an open-access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
repository_type Open Access Journal
institution_category Foreign Institution
institution US National Center for Biotechnology Information
building NCBI PubMed
collection Online Access
language English
format Online
author Feng, Wanyou
Chen, Shibei
Do, Dagiang
Liu, Qinyou
Deng, Yanfei
Lei, Xiaocan
Luo, Chan
Huang, Ben
Shi, Deshun
spellingShingle Feng, Wanyou
Chen, Shibei
Do, Dagiang
Liu, Qinyou
Deng, Yanfei
Lei, Xiaocan
Luo, Chan
Huang, Ben
Shi, Deshun
Isolation and Identification of Prepubertal Buffalo (Bubalus bubalis) Spermatogonial Stem Cells
author_facet Feng, Wanyou
Chen, Shibei
Do, Dagiang
Liu, Qinyou
Deng, Yanfei
Lei, Xiaocan
Luo, Chan
Huang, Ben
Shi, Deshun
author_sort Feng, Wanyou
title Isolation and Identification of Prepubertal Buffalo (Bubalus bubalis) Spermatogonial Stem Cells
title_short Isolation and Identification of Prepubertal Buffalo (Bubalus bubalis) Spermatogonial Stem Cells
title_full Isolation and Identification of Prepubertal Buffalo (Bubalus bubalis) Spermatogonial Stem Cells
title_fullStr Isolation and Identification of Prepubertal Buffalo (Bubalus bubalis) Spermatogonial Stem Cells
title_full_unstemmed Isolation and Identification of Prepubertal Buffalo (Bubalus bubalis) Spermatogonial Stem Cells
title_sort isolation and identification of prepubertal buffalo (bubalus bubalis) spermatogonial stem cells
description 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.
publisher Asian-Australasian Association of Animal Production Societies (AAAP) and Korean Society of Animal Science and Technology (KSAST)
publishDate 2016
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5003965/
_version_ 1613638579009880064

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