Growth factors improve the proliferation of Jeju black pig muscle cells by regulating myogenic differentiation 1 and growth-related genes
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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 1.5×10<sup>5</sup> cells per well and cultured for 3 days. Western blot and quantitative real-time polymerase chain reaction were applied to verify the myogenic differentiation 1 (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 paired box 7. The proliferation rate of these muscle cells was significantly higher in a culture medium containing bFGF and epidermal growth factor + basic fibroblast growth factor (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 collagen type XXI alpha 1 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.
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 1.5×105 cells per well and cultured for 3 days. Western blot and quantitative real-time polymerase chain reaction were applied to verify the myogenic differentiation 1 (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 paired box 7. The proliferation rate of these muscle cells was significantly higher in a culture medium containing bFGF and epidermal growth factor + basic fibroblast growth factor (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 collagen type XXI alpha 1 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.
참고문헌 (42)
- Ghosh M, "The first comprehensive description of the expression profile of genes involved in differential body growth and the immune system of the Jeju Native Pig and miniature pig", Amino Acids, 51, pp.495~511 (2019)[DOI]
- Lunney JK, "Advances in swine biomedical model genomics", Int J Biol Sci, 3, pp.179~184 (2007)[DOI]
- In-Cheol Cho, "The Robust Phylogeny of Korean Wild Boar (Sus scrofa coreanus) Using Partial D-Loop Sequence of mtDNA", Molecules and Cells, 28(5), pp.423~430 (2009), 한국분자세포생물학회
- Ghosh M, "An integrated in silico approach for functional and structural impact of non-synonymous SNPs in the MYH1 gene in Jeju Native Pigs", BMC Genet, 17, pp.35~ (2016)[DOI]
- Sodhi SS, "Comparative transcriptomic analysis by RNA-seq to discern differential expression of genes in liver and muscle tissues of adult Berkshire and Jeju Native Pig", Gene, 546, pp.233~242 (2014)[DOI]
- 김계웅, "제주재래돼지의 성별 및 도체등급에 따른 도체특성 분석", 한국축산학회지, 52(4), pp.313~318 (2010), 한국축산학회
- Post MJ, "Cultured meat from stem cells : challenges and prospects", Meat Sci, 92, pp.297~301 (2012)[DOI]
- Musarò A, "Mouse cell culture", pp.101~111 (2010), Humana Press
- Yucel N, "Principles of Regenerative Medicine", pp.273~293 (2019), Academic Press
- Ben-Arye T, "Tissue engineering for clean meat production", Front Sustain Food Syst, 3, pp.46~ (2019)[DOI]
- Tidball JG, "Regulatory interactions between muscle and the immune system during muscle regeneration", Am J Physiol Regul Integr Comp Physiol, 298, pp.R1173~R1187 (2010)[DOI]
- Hernández-Hernández JM, "The myogenic regulatory factors, determinants of muscle development, cell identity and regeneration", Semin Cell Dev Biol, 72, pp.10~18 (2017)[DOI]
- Berkes CA, "MyoD and the transcriptional control of myogenesis", Semin Cell Dev Biol, 16, pp.585~595 (2005)[DOI]
- Blais A, "An initial blueprint for myogenic differentiation", Gene Dev, 19, pp.553~569 (2005)[DOI]
- Cao Y, "Genome-wide MyoD binding in skeletal muscle cells : a potential for broad cellular reprogramming", Dev Cell, 18, pp.662~674 (2010)[DOI]
- Soleimani VD, "Snail regulates MyoD binding-site occupancy to direct enhancer switching and differentiation-specific transcription in myogenesis", Mol Cell, 47, pp.457~468 (2012)[DOI]
- Yang J, "Isolation, culture and biological characteristics of multipotent porcine skeletal muscle satellite cells", Cell Tissue Bank, 18, pp.513~525 (2017)[DOI]
- Andres-Mateos E, "Impaired skeletal muscle regeneration in the absence of fibrosis during hibernation in 13-lined ground squirrels", PLoS One, 7, pp.e48884~ (2012)[DOI]
- Trippel SB, "Growth factors as therapeutic agents", Instr Course Lect, 46, pp.473~476 (1997)[DOI]
- Grounds M, "Towards understanding skeletal muscle regeneration", Pathol Res Pract, 187, pp.1~22 (1991)[DOI]
- Menetrey J, "Growth factors improve muscle healing in vivo", J Bone Joint Surg Br, 82, pp.131~137 (2000)[DOI]
- Xu C, "A zebrafish embryo culture system defines factors that promote vertebrate myogenesis across species", Cell, 155, pp.909~921 (2013)[DOI]
- Aghila Rani KG, "Effects of epidermal growth factor on proliferation and migration of cardiosphere-derived cells expanded from adult human heart", Growth Factors, 28, pp.157~165 (2010)[DOI]
- Braun T, "Transcriptional mechanisms regulating skeletal muscle differentiation, growth and homeostasis", Nat Rev Mol Cell Biol, 12, pp.349~361 (2011)[DOI]
- Tehrani HJ, "Effect of dexamethasone, insulin and EGF on the myogenic potential on human endometrial stem cell", Iran J Pharm Sci, 13, pp.659~664 (2014)
- Pirskanen A, "IGFs, insulin, Shh, bFGF, and TGF-β1 interact synergistically to promote somite myogenesis in vitro", Dev Biol, 224, pp.189~203 (2000)[DOI]
- Hongxue Shi, "Myoprotective effects of bFGF on skeletal muscle injury in pressure-related deep tissue injury in rats", Burns & Trauma, 4 (2016), Oxford University Press (OUP)[DOI]
- Liu X, "Proliferative effect of basic fibroblast growth factor and epidermal growth factor on muscle derived stem cells", Chinese J Repar Reconstr Surg, 20, pp.936~939 (2006)
- Nain Z, "Current understanding on tail regeneration in green anoles(Anolis carolinensis)", Cell Biol, 4, pp.9~17 (2016)[DOI]
- Palade J, "Identification of satellite cells from anole lizard skeletal muscle and demonstration of expanded musculoskeletal potential", Dev Biol, 433, pp.344~356 (2018)[DOI]
- Liang P, "CRISPR", Cas9-mediated gene editing in human tripronuclear zygotes, 2015, pp.6, 363~372
- Sun N, "Seamless correction of the sickle cell disease mutation of the HBB gene in human induced pluripotent stem cells using TALENs", Biotechnol Bioeng, 111, pp.1048~1053 (2014)[DOI]
- Maman S, "The beta subunit of hemoglobin(HBB2", HBB)suppresses neuroblastoma growth and metastasis, 2017, pp.77, 14~26
- Sato C, "Relationship between ST8SIA2, polysialic acid and its binding molecules, and psychiatric disorders", Biochim Biophys Acta Gen Subj, 1860, pp.1739~1752 (2016)[DOI]
- Shaw AD, "Characterisation of genetic variation in ST8SIA2 and its interaction region in NCAM1in patients with bipolar disorder", PLoS one, 9, pp.e92556~ (2014)[DOI]
- Brocco M, "Differential regulation of polysialyltransferase expression during hippocampus development : implications for neuronal survival", J Neurosci Res, 74, pp.744~753 (2003)[DOI]
- Hane M, "Effects of intronic single nucleotide polymorphisms(iSNPs)of a polysialyltransferase, ST8SIA2gene found in psychiatric disorders on its gene products", Biochem Biophys Res Commun, 478, pp.1123~1129 (2016)[DOI]
- Goetsch SC, "Transcriptional profiling and regulation of the extracellular matrix during muscle regeneration", Physiol Genomics, 14, pp.261~271 (2003)[DOI]
- Bonnet N, "Periostin action in bone", Mol Cell Endocrinol, 432, pp.75~82 (2016)[DOI]
- Fitzgerald J, "A new FACIT of the collagen family : COL21A1", FEBS Lett, 505, pp.275~280 (2001)[DOI]
- Chou M-Y, "Genomic organization and characterization of the human type XXI collagen(COL21A1)gene", Genomics, 79, pp.395~401 (2002)[DOI]
- Nihashi Y, "Distinct cell proliferation, myogenic differentiation, and gene expression in skeletal muscle myoblasts of layer and broiler chickens", Sci Rep, 9, pp.16527~ (2019)[DOI]