Immortalization of bovine germ line stem cells by c-myc and hTERT
| Autor: | Li Wang, Shi-Qiang Zhang, Yong Zhang, Zhi-Xing An, Shu-Ying Peng, Cong-Ming Bi, Ning Lv, Aladaer Qi |
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| Rok vydání: | 2007 |
| Předmět: |
Telomerase
Time Factors Transgene Biology Germline Cell Line Proto-Oncogene Proteins c-myc Endocrinology Food Animals Animals Telomerase reverse transcriptase RNA Messenger Cell Proliferation Expression vector Reverse Transcriptase Polymerase Chain Reaction General Medicine Transfection Immunohistochemistry Molecular biology Germ Cells Gene Expression Regulation Cell culture Cattle Animal Science and Zoology Stem cell Genetic Engineering |
| Zdroj: | Animal Reproduction Science. 100:371-378 |
| ISSN: | 0378-4320 |
| DOI: | 10.1016/j.anireprosci.2006.10.017 |
| Popis: | 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 GFRα-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. |
| Databáze: | OpenAIRE |
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