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Lentiviral vector transfection of chicken embryo primordial germ cells in vitro

Published online by Cambridge University Press:  27 June 2008

Ding Hong-Mei
Affiliation:
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
Xu Shi-Yong
Affiliation:
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
Shao Gen-Bao
Affiliation:
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
Sun Yan
Affiliation:
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
Wang Meng
Affiliation:
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
Liu Hong-Lin**
Affiliation:
College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China
*
*Corresponding author. E-mail: liuhonglin@263.net

Abstract

Lentiviruses as gene transfer vectors have been used successfully to transfect mammal embryonic stem cells and germline stem cells, but this has not been attempted in avian primordial germ cells (PGCs). PGCs were isolated from the gonads of Isa-brown chicken embryos at stage 28 and co-cultured with gonadal stroma cells. A lentiviral vector pLenti-CMV-EGFP was constructed and the virus harvested by cotransfecting 293FT cells with the vector and packaging plasmids. Concentrated lentiviruses were used to transfect chicken PGCs, the transfection efficiency was up to 24.19%.

Type
Research Papers
Copyright
Copyright © China Agricultural University 2008

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Footnotes

First published in Journal of Agricultural Biotechnology 2007, 15(4): 612–616

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