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Immortalization of primary sheep embryo kidney cells

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Abstract

Sheep primary epithelial cells are short-lived in cell culture systems. For long-term in vitro studies, primary cells need to be immortalized. This study aims to establish and characterize T immortalized sheep embryo kidney cells (TISEKC). In this study, we used fetal lamb kidneys to derive primary cultures of epithelial cells. We subsequently immortalized these cells using the large T SV40 antigen to generate crude TISEKC and isolate TISEKC clones. Among numerous clones of immortalized cells, the selected TISEKC-5 maintained active division and cell growth over 20 passages but lacked expression of the oncogenic large T SV40 antigen. Morphologically, TISEKC-5 maintained their epithelial aspect similar to the parental primary epithelial cells. However, their growth properties showed quite different patterns. Crude TISEKC, as well as the clones of TISEKC proliferated highly in culture compared to the parental primary cells. In the early passages, immortalized cells showed heterogeneous polyploidy but in the late passages the karyotype of immortalized cells became progressively stable, identical to that of the primary cells, because the TISEKC-5 cell line has lost the large SV40 T antigen expression, this cell line is a valuable tool for veterinary sciences and biotechnological productions.

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Acknowledgments

The authors wish to gratefully thank Myriam Khattabi and Marcelline Samadane for excellent technical help.

Funding

This work was supported by the Algerian Ministry of High Education and Scientific Research, the University of Sciences and Technology “Houari Boumediene” Algiers, the French INRA, and the University of Grenoble Alps.

Author information

Authors and Affiliations

  1. Laboratory of Molecular and Cellular Biology, Unit of Genetics, Faculty of Biological Sciences, University of Sciences and Technology “Houari Boumediene”, Algiers, Algeria

    N. Seridi

  2. Laboratory of Production and Development of Viral Veterinary Vaccines, Pasteur Institute of Algeria, Algiers, Algeria

    M. Hamidouche & N. Belmessabih

  3. INRAE/UGA USC 1450, Pathogenesis and Lentivirus Vaccination Laboratory, PAVAL Lab, Université Grenoble Alpes, 38041, Grenoble Cedex 9, France

    S. El Kennani, J. Gagnon & Y. Chebloune

  4. CHU Grenoble Alpes, UM de Génétique Chromosomique, Grenoble, France

    G. Martinez & C. Coutton

  5. INSERM U1209, CNRS UMR 5309, Institute for Advanced Biosciences, Université Grenoble Alpes, 38000, Grenoble, France

    G. Martinez & C. Coutton

  6. VetAgro Sup, UPSP ICE 2011.03.101, Laboratoire d’Histopathologie, Université de Lyon, Marcy-l’Etoile, France

    T. Marchal

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  1. N. Seridi
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  2. M. Hamidouche
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  3. N. Belmessabih
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  4. S. El Kennani
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  5. J. Gagnon
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  6. G. Martinez
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  7. C. Coutton
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  8. T. Marchal
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  9. Y. Chebloune
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Corresponding author

Correspondence to Y. Chebloune.

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Editor: Tetsuji Okamoto

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Seridi, N., Hamidouche, M., Belmessabih, N. et al. Immortalization of primary sheep embryo kidney cells. In Vitro Cell.Dev.Biol.-Animal 57, 76–85 (2021). https://doi.org/10.1007/s11626-020-00520-y

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  • DOI: https://doi.org/10.1007/s11626-020-00520-y

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