title: Immortalization of bovine germ line stem cells by c-myc and hTERT.
abstract: The limited life span of bovine germ line stem cells in vitro is one of the obstacles to spermatogenesis analysis, genetic manipulation and generating transgenic animal. The aim of this study is to establish immortalized bovine germ line stem cells by c-myc or hTERT. We constructed pEMY and pETE expression vectors and transfected germ line cells from 5-month-old bovine. After G418 screening, four types of positive clones were obtained. The results showed that they expressed exogenous genes c-myc or hTERT at mRNA and protein level by RT-PCR and Western blotting detection. Presumable cell lines GM7, GT3, GMT5 all expressed germ-line-stem-cell-specific makers by immunocytochemical analysis, such as c-kit, Oct-4 and GFRalpha-1. The putative cell lines also had higher capacity of proliferation than freshly isolated bovine spermatogonial stem cells. So we can conclude that exogenous genes c-myc or hTERT have integrated into the genome of bovine germ cells and upregulated the expression of telomerase.
authors.affiliation: Institute of Bio-engineering, Northwest Sci-Tech University of Agriculture and Forestry, No. 3 TaiCheng Road, Yangling 712100, PR China.
authors.forename: Cong-Ming
authors.initials: CM
authors.lastname: Bi
authors.forename: Shi-Qiang
authors.initials: SQ
authors.lastname: Zhang
authors.forename: Yong
authors.initials: Y
authors.lastname: Zhang
authors.forename: Shu-Ying
authors.initials: SY
authors.lastname: Peng
authors.forename: Li
authors.initials: L
authors.lastname: Wang
authors.forename: Zhi-Xing
authors.initials: ZX
authors.lastname: An
authors.forename: Aladaer
authors.initials: A
authors.lastname: Qi
authors.forename: Ning
authors.initials: N
authors.lastname: Lv
mesh_terms: D000818:Animals; D002417:Cattle / Q000235:genetics*; D002460:Cell Line; D049109:Cell Proliferation; D005786:Gene Expression Regulation; D005818:Genetic Engineering; D005854:Germ Cells / Q000166:cytology* / Q000378:metabolism*; D007150:Immunohistochemistry; D016271:Proto-Oncogene Proteins c-myc / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary; D019098:Telomerase / Q000235:genetics* / Q000378:metabolism; D013997:Time Factors
chemical_list: D016271:Proto-Oncogene Proteins c-myc; D012333:RNA, Messenger; C509186:TERT protein, human; D019098:Telomerase