Acta Veterinaria et Zootechnica Sinica ›› 2021, Vol. 52 ›› Issue (3): 676-682.doi: 10.11843/j.issn.0366-6964.2021.03.011

• ANIMAL GENETICS AND BREEDING • Previous Articles     Next Articles

Isolation, Culture and Identification of Muscle Satellite Cells of Chicken

DAI Wei1,2,3, SONG Ruilong1,2,3, ZHANG Yuanhao1,2,3, ZOU Hui1,2,3, GU Jianhong1,2,3, YUAN Yan1,2,3, BIAN Jianchun1,2,3, LIU Xuezhong1,2,3*   

  1. 1. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China;
    2. Jiangsu Co-innovation Center for Prevention and Control of Important Animal Infectious Diseases and Zoonoses, Yangzhou 225009, China;
    3. Joint International Research Laboratory of Agriculture and Agri-Product Safety of the Ministry of Education of China, Yangzhou University, Yangzhou 225009, China
  • Received:2020-08-06 Online:2021-03-23 Published:2021-03-24

Abstract: The purpose of the study was to establish methods for isolation, culture, purification, differentiation and identification of chicken skeletal muscle satellite cells (MSC) in vitro. Selecting SPF chicken embryos of 12 embryonic age, integrating the principles of loose muscle fiber and free fiber matrix layer cells, the enzymes used to separate cells, cell purification and differentiation induction methods were optimized, and the MSC was identified from the aspects of morphology, cell differentiation ability, growth curve, immunofluorescence detection, RT-PCR identification, etc. Therefore, a method of digesting, separating and culturing chicken skeletal muscle satellite cells with mixed enzymes was proposed. The results showed that the separated and purified cells had strong refractive index and adhered to the wall after 24 hours, showing a spindle shape; after induced differentiation, it could form neatly arranged multinucleated myotubes; the cell growth curve measured by using CCK-8 method showed a typical "S" shape; after the optimization, the cell survival rate was (90.82±1.294)%, the cell purity was (90.44±1.264)%; immunofluorescence detection of the marker genes Pax7 and Desmin were positive; RT-PCR detection of the marker genes Pax7, MyHC, MyoD1 were positive; and the expression of the marker gene Pax7 before differentiation was 1.705 times that after differentiation, and the expression of the marker gene MyHC after differentiation was 13.073 times that before differentiation. This study established a quick and simple method for culturing chicken skeletal muscle satellite cells in vitro, which provides a good cell model for studying the biological mechanism of chicken skeletal muscle cells, optimization of broiler breeds, and cell transplantation repair.

Key words: muscle satellite cell, isolation, culture, induced differentiation, identification

CLC Number: