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DOI: https://doi.org/10.5713/ab.20.0585    [Accepted] Published online January 1, 2021.
Growth factors improve the proliferation of Jeju black pig muscle cells by regulating MyoD and growth-related genes
Jinryong Park1  , Jeongeun Lee1  , Ki-Duk Song2,7  , Sung-Jo Kim3  , Dae Cheol Kim4  , Sang Cheol Lee5  , Young June Son5  , Hyun Woo Choi6,7,*  , Kwanseob Shim1,7,* 
1Department of Animal Biotechnology, Jeonbuk National University, Jeonju 54896, Korea
2The Animal Molecular Genetics and Breeding Center, Jeonbuk National University, Jeonju 54896, Korea
3Division of Cosmetics and Biotechnology, Hoseo University, Asan 31499, Korea
4Livestock Promotion Institute, Jeju Special Self-Governing Province, Jeju 63122, Korea
5Cronex Inc, Cheongju 28174, Korea
6Department of Animal Science, Jeonbuk National University, Jeonju 54896, Korea
7Department of Agricultural Convergence Technology, Jeonbuk National University, Jeonju 54896, Korea
Correspondence:  Hyun Woo Choi,Email: choihw@jbnu.ac.kr
Kwanseob Shim, Tel: +82-63-270-2609, Fax: +82-63-238-2614, Email: ksshim@jbnu.ac.kr
Received: 21 August 2020   • Revised: 16 November 2020   • Accepted: 12 December 2020
Abstract
Objective
The growth rate of pigs is related to differentiation and proliferation of muscle cells, which are regulated by growth factors and expression of growth-related genes. Thus, the objective of this study was to establish optimal culture conditions for Jeju black pig (JBP) muscle cells and determine the relationship of various factors involved in muscle growth with the proliferation of JBP muscle cells.
Methods
Muscles were taken from the femur skeletal muscle of JBP embryos. After isolation of the muscle cells, cells were cultured in a 6-well plate under four different culture conditions to optimize culture conditions for JBP muscle cells. To analyze proliferation rate of JBP muscle cells, these muscle cells were seeded into 6-well plates at a density of 1.5 x 105 cells per well and cultured for 3 days. Western blot and quantitative real-time polymerase chain reaction were applied to verify the MyoD expression and growth-related gene expression in JBP muscle cells, respectively.
Results
We established a muscle cell line from JBP embryos and optimized its culture conditions. These muscle cells were positive for MyoD, but not for Pax7. The proliferation rate of these muscle cells was significantly higher in a culture medium containing bFGF and EGF+bFGF than that without a growth factor or containing EGF alone. Treatment with EGF and bFGF significantly induced the expression of MyoD protein, an important transcription factor in muscle cells. Moreover, we checked the changes of expression of growth-related genes in JBP muscle cells by presence or absence of growth factors. Expression level of COL21A1 gene was changed only when EGF and bFGF were added together to culture media for JBP muscle cells.
Conclusion
Concurrent use of EGF and bFGF increased the expression of MyoD protein, thus regulating the proliferation of JBP muscle cells and the expression of growth-related genes.
Keywords: Pig; Muscle Cell; Growth Factor; MyoD; Growth-related Gene
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