Hostname: page-component-848d4c4894-hfldf Total loading time: 0 Render date: 2024-06-01T01:32:53.966Z Has data issue: false hasContentIssue false
  • English
  • Français

Lentiviral vector transfection of chicken embryo primordial germ cells in vitro

Published online by Cambridge University Press:  27 June 2008

Ding Hong-Mei ,
Xu Shi-Yong ,
Shao Gen-Bao ,
Sun Yan ,
Wang Meng  and
Liu Hong-Lin*
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
Get access Rights & Permissions [Opens in a new window]

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%.

Keywords

chicken embryoslentivirusprimordial germ cellstransfection efficiency
Type
Research Papers
Information
Chinese Journal of Agricultural Biotechnology , Volume 5 , Issue 1 , April 2008 , pp. 1 - 5
Copyright
Copyright © China Agricultural University 2008

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

Footnotes

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

References

Allioli, N, Thomas, JL, Chebloune, Y, Nigon, VM, Verdier, G and Legras, C (1994) Use of retroviral vectors to introduce and express the beta-galactosidase marker gene in cultured chicken primordial germ cells. Developmental Biology 165: 30–37.CrossRefGoogle ScholarPubMed
Chang, I, Tajima, A, Chikamune, T and Ohno, T (1995) Proliferation of chick primordial germ cells cultured on stroma cells from the germinal ridge. Cell Biology International 19(2): 143–149.CrossRefGoogle ScholarPubMed
Hamra, FK, Gatlin, J, Chapman, KM, et al. (2002) Production of transgenic rats by lentiviral transduction of male germ-line stem cells. Proceedings of the National Academy of Sciences of the USA 99(23): 14931–14936.CrossRefGoogle ScholarPubMed
Han, JY, Shoffner, RN and Guise, KS (1994) Gene transfer by manipulation of primordial germ cells in the chicken. Asian–Australian Journal Animal Science 7(3): 427–434.CrossRefGoogle Scholar
Jeong, DK, Hong, YH and Han, JY (2002) Simple separation of chicken gonadal primordial germ cells with and without foreign genes. Cell Biology International 26(7): 647–651.CrossRefGoogle ScholarPubMed
McGrew, MJ, Sherman, A, Ellard, FM, et al. (2004) Efficient production of germline transgenic chickens using lentiviral vectors. European Molecular Biology Organization 5(7): 728–733.Google ScholarPubMed
Nagano, M, Watson, DJ, Ryu, BY, Wolfe, JH and Brinster, RL (2002) Lentiviral vector transduction of male germ line stem cells in mice. Federation of European Biochemical Societies 524: 111–115.CrossRefGoogle ScholarPubMed
Nakamura, M, Kuwana, T, Miyayama, Y, Fujimoto, T (1988) Extragonadal distribution of primordial germ cells in the early chick embryo. Anatomical Record 222(1): 90–94.CrossRefGoogle ScholarPubMed
Park, TS and Han, JY (2000) Derivation and characterization of pluripotent embryonic germ cells in chicken. Molecular Reproduction and Development 56: 475–482.3.0.CO;2-M>CrossRefGoogle ScholarPubMed
Park, TS, Jeong, DK, Kim, JN, et al. (2003) Improved germline transmission in chicken chimeras produced by transplantation of gonadal primordial germ cells into recipient embryos. Biology of Reproduction 68: 1657–1662.CrossRefGoogle ScholarPubMed
Petitte, JN, Liu, G and Yang, Z (2004) Avian pluripotent stem cells. Mechanisms of Development 121: 1159–1168.CrossRefGoogle ScholarPubMed
Pfeifer, A, Lkawa, M, Dayn, Y and Verma, IM (2002) Transgenesis by lentiviral vectors: lack of gene silencing in mammalian embryonic stem cells and preimplantation embryos. Proceedings of the National Academy of Sciences of the USA 99(4):2140–2145.CrossRefGoogle ScholarPubMed
Sang, H (2006) Transgenesis sunny-side up. Nature Biotechnology 24(8): 955–956.CrossRefGoogle ScholarPubMed
Wei, Q, Croy, BA and Etches, RJ (2001) Selection of genetically modified chicken blastodermal cells by magnetic-activated cell sorting. Poultry Science 80: 1671–1678.CrossRefGoogle ScholarPubMed