[논문]Serum-free cultures of C2C12 cells show different muscle phenotypes which can be estimated by metabolic profiling

Jang, Mi; Scheffold, Jana; Røst, Lisa Marie; Cheon, Hyejeong; Bruheim, Per

doi:10.1038/s41598-022-04804-z

Serum-free cultures of C2C12 cells show different muscle phenotypes which can be estimated by metabolic profiling 원문보기

Scientific reports, v.12, 2022년, pp.827 -

Jang, Mi (Department of Biotechnology and Food Science, Norwegian University of Science and Technology, Hogskoleringen 1, 7491 Trondheim, Norway) , Scheffold, Jana (Department of Biotechnology and Food Science, Norwegian University of Science and Technology, Hogskoleringen 1, 7491 Trondheim, Norway) , Røst, Lisa Marie (Department of Biotechnology and Food Science, Norwegian University of Science and Technology, Hogskoleringen 1, 7491 Trondheim, Norway) , Cheon, Hyejeong (PoreLab, Department of Physics, Norwegian University of Science and Technology, Hogskoleringen 1, 7491 Trondheim, Norway) , Bruheim, Per (Department of Biotechnology and Food Science, Norwegian University of Science and Technology, Hogskoleringen 1, 7491 Trondheim, Norway)

Abstract ▼ AI-Helper

In vitro skeletal muscle cell production is emerging in the field of artificial lab-grown meat as alternative future food. Currently, there is an urgent paradigm shift towards a serum replacement culture system. Surprisingly, little is known about the impact of serum-free culture on skeletal muscle cells to date. Therefore, we performed metabolic profiling of the C2C12 myoblasts and myotubes in serum-free mediums (B27, AIM-V) and compared it with conventional serum supplementation culture. Furthermore, cell morphology, viability, and myogenic differentiation were observed for 7 days of cultivation. Intriguingly, the metabolic difference is more dominant between the cell status than medium effects. In addition, proliferative myoblast showed more distinct metabolic differences than differentiated myotubes in different culture conditions. The intracellular levels of GL3P and UDP-GlcNAc were significantly increased in myotubes versus myoblast. Non-essential amino acids and pyruvate reduction and transamination showed significant differences among serum, B27, and AIM-V cultures. Intracellular metabolite profiles indicated that C2C12 myotubes cultured in serum and B27 had predominant glycolytic and oxidative metabolism, respectively, indicating fast and slow types of muscle confirmed by MHC immunostaining. This work might be helpful to understand the altered metabolism of skeletal muscle cells in serum-free culture and contribute to future artificial meat research work.

참고문헌 (59)

1. Price PJ Best practices for media selection for mammalian cells In Vitro Cell. Dev. Biol. Anim. 2017 53 673 681 28726187

상세보기
2. Brindley DA Peak serum: Implications of serum supply for cell therapy manufacturing Regen. Med. 2012 7 7 13 22168489
3. Ben-Arye T Levenberg S Tissue engineering for clean meat production Front. Sustain. Food Syst. 2019 3 66
4. van der Valk J Optimization of chemically defined cell culture media—Replacing fetal bovine serum in mammalian in vitro methods Toxicol. In Vitro 2010 24 1053 1063 20362047

상세보기
5. Liu CH Liu YX Wu WC Facile development of medium optimization for antibody production: Implementation in spinner flask and hollow fiber reactor Cytotechnology 2018 70 1631 1642 30284074

상세보기
6. McGillicuddy N Floris P Albrecht S Bones J Examining the sources of variability in cell culture media used for biopharmaceutical production Biotechnol. Lett. 2018 40 5 21 28940015

상세보기
7. Karnieli O A consensus introduction to serum replacements and serum-free media for cellular therapies Cytotherapy 2017 19 155 169 28017599

상세보기
8. Orellana N A new edible film to produce in vitro meat Foods 2020 9 185

상세보기
9. MacQueen LA Muscle tissue engineering in fibrous gelatin: implications for meat analogs npj Sci. Food 2019 3 1 12 31304273
10. Zhang G Challenges and possibilities for bio-manufacturing cultured meat Trends Food Sci. Technol. 2020 97 66
11. Kolkmann AM Post MJ Rutjens MAM van Essen ALM Moutsatsou P Serum-free media for the growth of primary bovine myoblasts Cytotechnology 2020 72 111 120 31884572

상세보기
12. Will K Kuzinski J Palin M-F Hildebrandt J-P Rehfeldt C A second look at leptin and adiponectin actions on the growth of primary porcine myoblasts under serum-free conditions Arch. Anim. Breed. 2014 57 1 10
13. Cai A Myogenic differentiation of primary myoblasts and mesenchymal stromal cells under serum-free conditions on PCL-collagen I-nanoscaffolds BMC Biotechnol. 2018 18 1 12 29316906
14. Bodiou V Moutsatsou P Post MJ Microcarriers for upscaling cultured meat production Front. Nutr. 2020 7 10 32154261

상세보기
15. Lawson MA Purslow PP Differentiation of myoblasts in serum-free media: Effects of modified media are cell line-specific Cells Tissues Organs 2000 167 130 137 10971037

상세보기
16. Molnar P Wang W Natarajan A Rumsey JW Hickman JJ Photolithographic patterning of C2C12 myotubes using vitronectin as growth substrate in serum-free medium Biotechnol. Prog. 2007 23 265 268 17269697

상세보기
17. Fujita H Endo A Shimizu K Nagamori E Evaluation of serum-free differentiation conditions for C2C12 myoblast cells assessed as to active tension generation capability Biotechnol. Bioeng. 2010 107 894 901 20635352

상세보기
18. Guo G Serum-based culture conditions provoke gene expression variability in mouse embryonic stem cells as revealed by single-cell analysis Cell Rep. 2016 14 956 965 26804902

상세보기
19. Luhur A Adapting drosophila melanogaster cell lines to serum-free culture conditions G3 Genes Genomes Genet. 2020 10 4541 4551
20. Guijas C Montenegro-Burke JR Warth B Spilker ME Siuzdak G Metabolomics activity screening for identifying metabolites that modulate phenotype Nat. Biotechnol. 2018 36 316 320 29621222

상세보기
21. Muroya S Ueda S Komatsu T Miyakawa T Ertbjerg P Meatabolomics: Muscle and meat metabolomics in domestic animals Metabolites 2020 10 66
22. Røst LM Absolute quantification of the central carbon metabolome in eight commonly applied prokaryotic and eukaryotic model systems Metabolites 2020 10 56
23. Kumar K Venkatraman V Bruheim P Adaptation of central metabolite pools to variations in growth rate and cultivation conditions in Saccharomyces cerevisiae Microb. Cell Factories 2021 20 1 16
24. Huang B Mdfi promotes C2C12 cell differentiation and positively modulates fast-to-slow-twitch muscle fiber transformation Front. Cell Dev. Biol. 2021 9 605875 33553177

상세보기
25. Daskalaki E Pillon NJ Krook A Wheelock CE Checa A The influence of culture media upon observed cell secretome metabolite profiles: The balance between cell viability and data interpretability Anal. Chim. Acta 2018 1037 338 350 30292310

상세보기
26. Hwang SY Folic acid promotes the myogenic differentiation of C2C12 murine myoblasts through the Akt signaling pathway Int. J. Mol. Med. 2015 36 1073 1080 26310574
27. Listrat A How muscle structure and composition influence meat and flesh quality Sci. World J. 2016 2016 66
28. Gupta SK Metabolic engineering of CHO cells for the development of a robust protein production platform PLoS ONE 2017 12 66
29. la Fuente IMD On the dynamics of the adenylate energy system: Homeorhesis vs homeostasis PLoS ONE 2014 9 108 676
30. Levitt DE Chalapati N Prendergast MJ Simon L Molina PE Ethanol-impaired myogenic differentiation is associated with decreased myoblast glycolytic function Alcohol. Clin. Exp. Res. 2020 44 2166 2176 32945016
31. Ryall JG Metabolic reprogramming as a novel regulator of skeletal muscle development and regeneration FEBS J. 2013 280 4004 4013 23402377

상세보기
32. Fortini P Iorio E Dogliotti E Isidoro C Coordinated metabolic changes and modulation of autophagy during myogenesis Front. Physiol. 2016 7 237 27378945

상세보기
33. Tomczak KK Expression profiling and identification of novel genes involved in myogenic differentiation FASEB J. 2004 18 403 405 14688207
34. Fortini P The fine tuning of metabolism, autophagy and differentiation during in vitro myogenesis Cell Death Dis. 2016 7 66
35. Homolak J Is galactose a hormetic sugar? Evidence from rat hippocampal redox regulatory network bioRxiv 2021 10.1101/2021.03.08.434370
36. Luo S Comparative sensitivity of proliferative and differentiated intestinal epithelial cells to the food contaminant, deoxynivalenol Environ. Pollut. 2021 277 116 818
37. Mugabo Y Identification of a mammalian glycerol-3-phosphate phosphatase: Role in metabolism and signaling in pancreatic β-cells and hepatocytes Proc. Natl. Acad. Sci. USA 2016 113 E430 E439 26755581
38. Kubišta V Accumulation of a stable phosphorus compound in glycolysing insect muscle Nature 1957 180 549 549 13477228

상세보기
39. Liu X Mitochondrial glycerol 3-phosphate dehydrogenase promotes skeletal muscle regeneration EMBO Mol. Med. 2018 10 66
40. Lambert M Bastide B Cieniewski-Bernard C Involvement of O-GlcNAcylation in the skeletal muscle physiology and physiopathology: Focus on muscle metabolism Front. Endocrinol. 2018 9 578

상세보기
41. Shi H Skeletal muscle O-GlcNAc transferase is important for muscle energy homeostasis and whole-body insulin sensitivity Mol. Metab. 2018 11 160 29525407

상세보기
42. Ogawa M Terminal differentiation program of skeletal myogenesis is negatively regulated by O-GlcNAc glycosylation Biochim. Biophys. Acta Gen. Subj. 2012 1820 24 32
43. Zumbaugh, M. D. Signaling Pathways Regulating Skeletal Muscle Metabolism and Growth (2021).
44. Kumar A Metabolomic analysis of primary human skeletal muscle cells during myogenic progression Sci. Rep. 2020 10 11824 32678274

상세보기
45. Gray LR Tompkins SC Taylor EB Regulation of pyruvate metabolism and human disease Cell. Mol. Life Sci. 2014 71 2577 24363178

상세보기
46. Hyatt JPK Muscle-specific myosin heavy chain shifts in response to a long-term high fat/high sugar diet and resveratrol treatment in nonhuman primates Front. Physiol. 2016 7 77 26973542

상세보기
47. Talbot J Maves L Skeletal muscle fiber type: using insights from muscle developmental biology to dissect targets for susceptibility and resistance to muscle disease Wiley Interdiscip. Rev. Dev. Biol. 2016 5 518 27199166
48. Abdelmoez AM Comparative profiling of skeletal muscle models reveals heterogeneity of transcriptome and metabolism Am. J. Physiol. Cell Physiol. 2020 318 C615 C626 31825657

상세보기
49. Brooks GA The science and translation of lactate shuttle theory Cell Metab. 2018 27 757 785 29617642
50. Holeček M Mičuda S Amino acid concentrations and protein metabolism of two types of rat skeletal muscle in postprandial state and after brief starvation Physiol. Res 2017 66 959 967 28937247
51. Mashima D Correlation between skeletal muscle fiber type and free amino acid levels in Japanese Black steers Anim. Sci. J. 2019 90 604 30811817
52. Komiya Y Correlation between skeletal muscle fiber type and responses of a taste sensing system in various beef samples Anim. Sci. J. 2020 91 e134 25
53. Roth, V. Doubling Time—Online computing with 2 points. Doubling Time Computing https://www.doubling-time.com/compute.php (2006).
54. Torres-Guzmán R Preclinical characterization of abemaciclib in hormone receptor positive breast cancer Oncotarget 2017 8 69493 69507 29050219

상세보기
55. Schindelin J Fiji: An open-source platform for biological-image analysis Nat. Methods 2012 9 676 682 22743772

상세보기
56. Søgaard CK “Two hits—one stone”; increased efficacy of cisplatin-based therapies by targeting PCNA’s role in both DNA repair and cellular signaling Oncotarget 2018 9 32448 30197755

상세보기
57. Stafsnes MH Røst LM Bruheim P Improved phosphometabolome profiling applying isotope dilution strategy and capillary ion chromatography-tandem mass spectrometry J. Chromatogr. B Anal. Technol. Biomed. Life Sci. 2018 1083 278 283
58. MetaboAnalyst. https://www.metaboanalyst.ca/ .
59. Droste P Miebach S Niedenführ S Wiechert W Nöh K Visualizing multi-omics data in metabolic networks with the software Omix—A case study Biosystems 2011 105 154 161 21575673

표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

내보내기 구분	파일저장 인쇄 메일전송
구성항목	기본정보 상세정보 관리번호, 논문명, 저널/프로시딩명, 저자 , 발행년, 권, 호, 시작페이지, 끝페이지, 발행기관 관리번호, 논문명, 대등논문명, 저자 , 저널/프로시딩명, 발행기관, 발행년, 발행언어, 권, 호, 시작페이지, 끝페이지, ISBN, ISSN, 주제분야, 키워드, 초록(한글), 초록(영문), 저자(소속기관)
저장형식	Text(ASCII format) Excel format RefWorks Direct Export RIS format (for Reference Manager, ProCite, EndNote), Scholar's Aids, Mendeley
메일정보	받는사람 (필수) @ 보내는사람 (선택) @ 제목 내용 KISTI 검색결과 이메일 서비스
안내	총 건의 자료가 검색되었습니다. 다운받으실 자료의 인덱스를 입력하세요. (1-10,000) 검색결과의 순서대로 최대 10,000건 까지 다운로드가 가능합니다. 데이타가 많을 경우 속도가 느려질 수 있습니다.(최대 2~3분 소요) 다운로드 파일은 UTF-8 형태로 저장됩니다. 파일의 내용이 제대로 보이지 않을실 때는 웹브라우저 상단의 보기 -> 인코딩 -> 자동선택 여부를 확인하십시오. ~ Text(ASCII format) Excel format

연합인증

Serum-free cultures of C2C12 cells show different muscle phenotypes which can be estimated by metabolic profiling 원문보기

Abstract ▼ AI-Helper

참고문헌 (59)

이 논문을 인용한 문헌

관련 콘텐츠

원문 보기

원문 URL 링크

오픈액세스(OA) 유형

이 논문과 함께 이용한 콘텐츠

AI-Helper ※ AI-Helper는 오픈소스 모델을 사용합니다.

선택된 텍스트