Skip to main content
SpringerLink
Log in

Isolation of chicken embryonic stem cell and preparation of chicken chimeric model

  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Chicken embryonic stem cells (ESCs) were separated from blastoderms at stage-X and cultured in vitro. Alkaline phosphatase activity and stage-specific embryonic antigen-1 staining was conducted to detect ESCs. Then, chicken ESCs were transfected with linearized plasmid pEGFP-N1 in order to produce chimeric chicken. Firstly, the optimal electrotransfection condition was compared; the results showed the highest transfection efficiency was obtained when the field strength and pulse duration was 280 V and 75 μs, respectively. Secondly, the hatchability of shedding methods, drilling a window at the blunt end of egg and drilling a window at the lateral shell of egg was compared, the results showed that the hatchability was the highest for drilling a window at the lateral shell of egg. Thirdly, the hatchability of microinjection (ESCs was microinjected into chick embryo cavity) was compared too, the results showed there were significant difference between the injection group transfected with ESCs and that of other two groups. In addition, five chimeric chickens were obtained in this study and EGFP gene was expressed in some organs, but only two chimeric chicken expressed EGFP gene in the gonad, indicating that the chimeric chicken could be obtained through chick embryo cavity injection by drilling a window at the lateral shell of egg.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5

Similar content being viewed by others

References

  1. Reedy SE, Leibo SP, Clark ME, Etches RJ (2007) Beyond freezing semen. Proceedings, first international symposium on the artificial insemination of poultry. Poultry Science Association, Savoy, pp 251–261

  2. Eyal-Giladi H, Kochav S (1976) From cleavage to primitive streak formation: a complementary normal table and a new look at the first stage of the development of the chick. I. General morphology. Dev Biol 49:321–337

    Article  PubMed  CAS  Google Scholar 

  3. Petitte JN, Clark ME, Liu G, Verrinder Gibbins AM, Etches RJ (1990) Production of somatic and germline chimeras in the chicken by transfer of early blastodermal cells. Development 108:185–189

    PubMed  CAS  Google Scholar 

  4. Bednarczyk M, Lakota P, Slomski R, Plawski A, Lipinski D, Siemieniako B, Lisowski M, Czekalski P, Grajewski B, Dluzniewska P (2002) Reconstitution of a chicken breed by inter semating of germline chimeric birds. Poult Sci 81:1347–1353

    PubMed  CAS  Google Scholar 

  5. Kino K, Pain B, Leibo SP, Cochran M, Clark ME, Etches RJ (1997) Production of chimeras from injection of frozen-thawed blastodermal cells. Poult Sci 76:753–760

    PubMed  CAS  Google Scholar 

  6. Sun M, Shi QQ, Yin YH, Fu DZ, Qin YR, Xu F, Li BC (2011) Generation of antiviral transgenic chicken using spermatogonial stem cell transfected in vivo. Afr J Biotechnol 10:15678–15683

    Article  CAS  Google Scholar 

  7. Wu XS, Wu H, Li BC, Zhou GY, Sun SY, Qin J, Chen GH, Musa HH (2009) Isolation, purification and culture of spermatogonia in chicken. J Anim Vet Adv 8:2418–2423

    Google Scholar 

  8. Jia YZ, Xiong YL, Liu GZ (2001) The culture embryo stem cells and preparation of the chimeric chicken. J Chin Exp Zool 11:93–94

    Google Scholar 

  9. Liu SC, Yan HF, Xiao BN (2001) The effect of donor blastoderm cells transferred into embryo for chicken embryonic development by eggs fenestration. Life Sci Res 5:270–274

    Google Scholar 

  10. Bednarczyk M, Lakota P, Siwek M (2000) Improvement of hatchability of chicken eggs injected by blastoderm cells. Poult Sci 79:1823–1828

    PubMed  CAS  Google Scholar 

  11. Tan XJ (2006) Transgenic chicken research mediated by blastocyst cells. China Agricultural University

  12. Thoraval P, Lasserre F, Coudert F (1994) Somatic and germline chicken chimeras obtained from Brown and White Leghorns by transfer of early blastodermal cells. Poult Sci 73:1897–1905

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The study was supported by Doctoral Program of Advanced Research Fund (20103250110006), “Talents in six field” funded projects of Jiangsu Province and National Nature Science Foundation of China (31272429).

Author information

Authors and Affiliations

  1. College of Animal Science and Technology, Yangzhou University, Yangzhou, 225009, People’s Republic of China

    Yani Zhang, Haiyan Yang, Zhentao Zhang, Qingqing Shi, Dan Wang, Mengmeng Zheng & Bichun Li

  2. Animal and Avian Science Department, University of Maryland, College Park, MD, 20742, USA

    Jiuzhou Song

Authors
  1. Yani Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Haiyan Yang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Zhentao Zhang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Qingqing Shi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Dan Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Mengmeng Zheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Bichun Li
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Jiuzhou Song
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Bichun Li.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Zhang, Y., Yang, H., Zhang, Z. et al. Isolation of chicken embryonic stem cell and preparation of chicken chimeric model. Mol Biol Rep 40, 2149–2156 (2013). https://doi.org/10.1007/s11033-012-2274-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-012-2274-8

Keywords

Search

Navigation