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Isolation, culture and identification of sheep skeletal muscle satellite cells

Author

Listed:
  • Sailuo Wan

    (College of Agricultural Engineering, Anhui Open University, Hefei, P.R. China
    College of Animal Science and Technology, Anhui Agricultural University, Hefei, P.R. China)

  • Qianqian Pan

    (College of Animal Science and Technology, Anhui Agricultural University, Hefei, P.R. China
    Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Anhui Agricultural University, Hefei, P.R. China)

  • Jinbo Wei

    (College of Animal Science and Technology, Anhui Agricultural University, Hefei, P.R. China
    Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Anhui Agricultural University, Hefei, P.R. China)

  • Cuiyun Zhu

    (College of Animal Science and Technology, Anhui Agricultural University, Hefei, P.R. China
    Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Anhui Agricultural University, Hefei, P.R. China)

  • Jing Jing

    (College of Animal Science and Technology, Anhui Agricultural University, Hefei, P.R. China
    Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Anhui Agricultural University, Hefei, P.R. China)

  • Shuaiqi Qin

    (College of Animal Science and Technology, Anhui Agricultural University, Hefei, P.R. China
    Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Anhui Agricultural University, Hefei, P.R. China)

  • Rongcui Hu

    (College of Animal Science and Technology, Anhui Agricultural University, Hefei, P.R. China
    Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Anhui Agricultural University, Hefei, P.R. China)

  • Mengyu Lou

    (College of Animal Science and Technology, Anhui Agricultural University, Hefei, P.R. China
    Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Anhui Agricultural University, Hefei, P.R. China)

  • Shuang Li

    (College of Animal Science and Technology, Anhui Agricultural University, Hefei, P.R. China
    Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Anhui Agricultural University, Hefei, P.R. China)

  • Yinghui Ling

    (College of Animal Science and Technology, Anhui Agricultural University, Hefei, P.R. China
    Anhui Province Key Laboratory of Local Livestock and Poultry Genetic Resource Conservation and Bio-Breeding, Anhui Agricultural University, Hefei, P.R. China)

Abstract

Skeletal muscle satellite cells (SMSCs) play an important role in muscle growth, regeneration and maintenance. This study aimed to isolate SMSCs from sheep, establish a system for isolation, culture and identification of SMSCs in vitro, and provide seed cells for subsequent studies. SMSCs were isolated and purified from newborn 2-day-old healthy sheep by collagenase type I and trypsin two-step digestion and pre-plating method. The results indicated that the isolated and purified SMSCs showed full spindle shape and strong refractive index. The cell growth curve detected by CCK-8 kit was typical "S" type. Immunofluorescence analysis showed that the isolated cells expressed SMSC marker proteins Pax7 and MyoD1. After induction of myogenic differentiation, the cells fused with each other to form multinucleated myotubes and expressed the myoblast specific marker MHC. RT-PCR results showed that the cells expressed SMSC marker gene Pax 7. This experiment established an in vitro isolation, purification and identification system for sheep skeletal muscle satellite cells, which provided a good cell model for studying the biological mechanism of sheep skeletal muscle cells, optimizing sheep breeds, and cell transplantation repair.

Suggested Citation

  • Sailuo Wan & Qianqian Pan & Jinbo Wei & Cuiyun Zhu & Jing Jing & Shuaiqi Qin & Rongcui Hu & Mengyu Lou & Shuang Li & Yinghui Ling, 2022. "Isolation, culture and identification of sheep skeletal muscle satellite cells," Czech Journal of Animal Science, Czech Academy of Agricultural Sciences, vol. 67(10), pages 416-423.
    • Handle: RePEc:caa:jnlcjs:v:67:y:2022:i:10:id:52-2022-cjas
    DOI: 10.17221/52/2022-CJAS
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    References listed on IDEAS

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    1. Marie-Claude Sincennes & Caroline E. Brun & Alexander Y. T. Lin & Tabitha Rosembert & David Datzkiw & John Saber & Hong Ming & Yoh-ichi Kawabe & Michael A. Rudnicki, 2021. "Acetylation of PAX7 controls muscle stem cell self-renewal and differentiation potential in mice," Nature Communications, Nature, vol. 12(1), pages 1-15, December.
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