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The fibronectin concentration that optimally maintains porcine satellite cells

  • Jae Ho Han (Department of Agricultural Convergence Technology, Jeonbuk National University) ;
  • Si Won Jang (Department of Agricultural Convergence Technology, Jeonbuk National University) ;
  • Ye Rim Kim (Department of Animal Science, Jeonbuk National University) ;
  • Hoon Jang (Department of Life Science, Jeonbuk National University) ;
  • Kwan Seob Shim (Department of Agricultural Convergence Technology, Jeonbuk National University) ;
  • Hyun Woo Choi (Department of Agricultural Convergence Technology, Jeonbuk National University)
  • Received : 2023.03.22
  • Accepted : 2023.06.26
  • Published : 2023.12.01

Abstract

Objective: 'Cultured meat' has been suggested as means of solving the problems associated with overpopulation and gas emissions. Satellite cells are a major component in the production of cultured meat; however, these cells cannot be maintained in vitro over long periods. Fibronectin is a glycoprotein that affects biological processes such as cell adhesion, differentiation, and migration. Unfortunately, the characteristics of porcine satellite cells grown in a long-term culture when exposed to fibronectin-coated dishes are unknown. The objective of this study was to investigate the appropriate concentration of fibronectin coated dishes for proliferation and maintenance of porcine satellite cells at long-term culture. Methods: In this study, we isolated the satellite cells and fibroblast cells with pre-plating method. We next analyzed the cell doubling time, cell cycle, and rate of expressed paired box 7 (Pax7) and myogenic differentiation 1 (MyoD1) in porcine satellite cells cultured with 20 ㎍/mL of fibronectin-, gelatin-, and non-coated dishes at early and late passage. We then analyzed the proliferation of porcine satellite cells with various concentrations of mixed gelatin/fibronectin. We next determined the optimal concentration of fibronectin that would encourage proliferation and maintenance of porcine satellite cells in a long-term culture. Results: Doubling time was lowest when 20 ㎍/mL of fibronectin was used (as tested during an early and late passage). Levels of expressed Pax7 and MyoD1, assessed using immunocytochemistry, were highest in cells grown using fibronectin-coated dishes. The proliferation of gelatin/fibronectin mixed coatings had no significant effect on porcine satellite cells. The concentration of 5 ㎍/mL fibronectin coated dishes showed the lowest doubling time and maintained expression of Pax7. Conclusion: Fibronectin with 5㎍/mL effectively maintains porcine satellite cells, a discovery that will be of interest to those developing the next generation of artificial meats.

Keywords

Acknowledgement

This work was supported by IPET through 'High Valueadded Food Technology Development Program', and was funded by the Ministry of Agriculture, Food and Rural Affairs (MAFRA) (322006-05-1-CG000).

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