[논문]Optimal Pre-Plating Method of Chicken Satellite Cells for Cultured Meat Production

Kim, So-Hee; Kim, Chan-Jin; Lee, Eun-Yeong; Son, Yu-Min; Hwang, Young-Hwa; Joo, Seon-Tea

doi:10.5851/kosfa.2022.e61

Optimal Pre-Plating Method of Chicken Satellite Cells for Cultured Meat Production 원문보기

Food science of animal resources, v.42 no.6, 2022년, pp.942 - 952

Kim, So-Hee (Division of Applied Life Science (BK21 Four), Gyeongsang National University) , Kim, Chan-Jin (Division of Applied Life Science (BK21 Four), Gyeongsang National University) , Lee, Eun-Yeong (Division of Applied Life Science (BK21 Four), Gyeongsang National University) , Son, Yu-Min (Division of Applied Life Science (BK21 Four), Gyeongsang National University) , Hwang, Young-Hwa (Institute of Agriculture & Life Science, Gyeongsang National University) , Joo, Seon-Tea (Division of Applied Life Science (BK21 Four), Gyeongsang National University)

Abstract ▼ AI-Helper

To establish a pre-plating method of chicken satellite cells with high purity, pre-plating was performed under culture conditions of 37℃ and 41℃, and the pre-plating time was set from a total of 3 hours to 6 hours in consideration of the cell attachment time. The purity of the cells was confirmed by staining paired box protein 7 (Pax7) after proliferation, and Pax7 expression was the highest in culture flasks shaken for 2 hours after incubation at 41℃ for 2 hours to prevent the attachment of satellite cells (p<0.05). Also, when pre-plating and proliferation were performed at 37℃ and 41℃, the Pax7 expression rate was higher at 41℃. The differentiation capabilities of the three groups (T3, T6, and T7) with high Pax7 expression were compared and the fusion index (%) and myotube formation area (%) determined by myosin heavy chain (MHC) staining was calculated. The T6 and T7 groups, which were cultured at 41℃, showed significantly higher values than the T3 group (p0.05). These results suggest that pre-plating at 41℃ for a total of 4 hours was the most efficient in terms of cost and time for purifying chicken satellite cells for cultured meat.

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표/그림 (6)

그림 Fig. 1. Pre-plating steps for chicken satellite cell purification.
그림 Fig. 2. Representative immunofluorescence images of pre-plated cells incubated for a set time. The cells were proliferated and immunostained with Hoechst 33342 (blue) and Pax7 (red). Scale bars=100 μm.
그림 Fig. 3. Percentage of Pax7-positive immuno-stained cells. Pax7+ (%) was calculated as the proportion of cells stained with Pax7 to Hoechst 33342. (A) Percentage of Pax7+ cells pre-plated at 37℃, (B) percentage of Pax7+ cells pre-plated at 41℃ (n=3; error bars=SD; p<0.05).
그림 Fig. 4. Pax7 expression of chicken satellite cells proliferated at 37℃ and 41℃. (A) Representative immunofluorescence images of cells pre-plated at a set time. The cells were proliferated and immunostained with Hoechst 33342 (blue) and Pax7 (red). Scale bars=100 μm. (B) Percentage of Pax7-positive cells in cells pre-plated under a set temperature. Pax7+ was calculated as the proportion of cells stained with Pax7 to Hoechst 33342 (n=3; error bars=SD; p<0.05).
그림 Fig. 5. Representative immunofluorescence images of differentiated cells pre-plated at different temperatures (37℃ and 41℃). The cells were immunostained with Hoechst 33342 (blue) and MHC (green). Scale bars=100 μm. MHC, myosin heavy chain.
그림 Fig. 6. Differentiation capacity of chicken satellite cells by temperature. (A) Fusion index (FI; %) of cells with high Pax7 expression. FI was calculated by adjusting the images in Fig. 4 using the ImageJ softwaare and ViaFuse macro program. FI, the number of nuclei inside myotubes. (B) Myotube formation area (MFA; %) in cells highly purified by pre-plating. MFA was calculated by the ImageJ software (n=3; error bars=SD; * p<0.05).

AI 본문요약
AI-Helper

데이터처리

Duncan’s multiple range test was performed to compare the means
The data are presented as the mean±SD and statistical significance was determined using one-way ANOVA and Student’s t-test

이론/모형

GNU-210308-C0028). All experimental steps complied with the IACUC standard procedures.
GNU-210308-C0028). All step of experiments were complied with IACUC standard procedure.

성능/효과

2)
Specifically, the most effective way to minimize fibroblast contamination was to pre-incubate the cell suspension for 2 h before the flask containing cells was shaken every 8 min for 2 h. Furthermore, satellite cells could differentiate and form myotubes better when chicken satellite cells were incubated at 41℃ from pre-plating through differentiation.
In the other six groups (T2–T7) with pre-plating, Pax7 expression was higher than in the T1 group, but it was not easy to compare them visually
Pax7 expression (%) was lower in the T2 group than in the T3 group (p<0.05) and expression in the T4 group was not significantly different from either the T2 or T3 group (p>0.05).
Pax7-positivity (%), which compared Pax7 expression to Hoechst staining, was significantly lower in the T1 group compared to the other groups because the satellite cell purity was only 45% (p<0.05; Fig
Satellite cell purity in the T2–T3 groups pre-plated at 37℃ was significantly higher than in the T1 group, and the T3 group showed the highest Pax7 expression (p<0.05)
The FI (%) was the highest in the T6 group cultured at 41℃ from pre-plating to differentiation, and there was no significant difference between the T6 and T7 groups (p>0.05; Fig
The T3 group showed significantly lower FI (%) values than the T6 and T7 groups (p<0.05)
The cells were stained with paired box protein 7 (Pax7) to confirm purification levels after proliferation, and myosin heavy chain (MHC) was stained to determine temperature suitability after differentiation. The immunofluorescence staining results were used to determine whether the pre-plating conditions were suitable for chicken satellite cell purification.
The myotube formation area (MFA; %) was also the highest in the T6 group among the three groups and the T3 group showed a relatively lower MFA (%) value than the T6 and T7 groups (p<0.05)
When pre-plating was conducted at different temperatures, as in the T3, T6, and T7 groups, which showed a high Pax7 expression (%) of more than 80%, cells were differentiated at each pre-plated temperature and stained with anti-MHC (Fig. 5).

참고문헌 (29)

Allouh MZ Yablonka-Reuveni Z Rosser BWC 2008 Pax7 reveals a greater frequency and concentration of satellite cells at the ends of growing skeletal muscle fibers J Histochem Cytochem 56 77 87 10.1369/jhc.7A7301.2007 17938281

상세보기
Bakhtiari N Hosseinkhani S Soleimani M Hemmati R Noori-Zadeh A Javan M Tashakor A 2016 Short-term ursolic acid promotes skeletal muscle rejuvenation through enhancing of SIRT1 expression and satellite cells proliferation Biomed Pharmacother 78 185 196 10.1016/j.biopha.2016.01.010 26898441

상세보기
Baquero-Perez B Kuchipudi SV Nelli RK Chang KC 2012 A simplified but robust method for the isolation of avian and mammalian muscle satellite cells BMC Cell Biol 13 16 10.1186/1471-2121-13-16 22720831

상세보기
Benedetti A Cera G De Meo D Villani C Bouche M Lozanoska-Ochser B 2021 A novel approach for the isolation and long-term expansion of pure satellite cells based on ice-cold treatment Skelet Muscle 11 7 10.1186/s13395-021-00261-w 33731194

상세보기
Bischoff R 1997 Chemotaxis of skeletal muscle satellite cells Dev Dyn 208 505 515 10.1002/(SICI)1097-0177(199704)208:4<505::AID-AJA6>3.0.CO;2-M 9097022

상세보기
Choi KH Kim M Yoon JW Jeong J Ryu M Jo C Lee CK 2020 Purification of pig muscle stem cells using magnetic-activated cell sorting (MACS) based on the expression of cluster of differentiation 29 (CD29) Food Sci Anim Resour 40 852 859 10.5851/kosfa.2020.e51 32968735

원문보기 상세보기
Clevers H 2015 What is an adult stem cell? Science 350 1319 1320 10.1126/science.aad7016 26659043

상세보기
Ding S Swennen GNM Messmer T Gagliardi M Molin DGM Li C Zhou G Post MJ 2018 Maintaining bovine satellite cells stemness through p38 pathway Sci Rep 8 10808 10.1038/s41598-018-28746-7 30018348

상세보기
Ding S Wang F Liu Y Li S Zhou G Hu P 2017 Characterization and isolation of highly purified porcine satellite cells Cell Death Discov 3 17003 10.1038/cddiscovery.2017.3 28417015

상세보기
Doumit ME Merkel RA 1992 Conditions for isolation and culture of porcine myogenic satellite cells Tissue Cell 24 253 262 10.1016/0040-8166(92)90098-R 1589873

상세보기
Guan X Zhou J Du G Chen J 2022 Bioprocessing technology of muscle stem cells: Implications for cultured meat Trends Biotechnol 40 721 734 10.1016/j.tibtech.2021.11.004 34887105

상세보기
Hartig SM 2013 Basic image analysis and manipulation in ImageJ Curr Protoc Mol Biol 102 14.15.1 14.15.12 10.1002/0471142727.mb1415s102 23547012

상세보기
Hinkle ER Essader TO Gentile GM Giudice J 2021 ViaFuse: Fiji macros to calculate skeletal muscle cell viability and fusion index Skeletal Muscle 11 28 10.1186/s13395-021-00284-3 34915930

상세보기
Ismail I Hwang YH Joo ST 2020 Meat analog as future food: A review J Anim Sci Technol 62 111 120 10.5187/jast.2020.62.2.111 32292920

원문보기 상세보기
Joo ST Choi JS Hur SJ Kim GD Kim CJ Lee EY Bakhsh A Hwang YH 2022 A comparative study on the taste characteristics of satellite cell cultured meat derived from chicken and cattle muscles Food Sci Anim Resour 42 175 185 10.5851/kosfa.2021.e72 35028582

원문보기 상세보기
Kadim IT Mahgoub O Baqir S Faye B Purchas R 2015 Cultured meat from muscle stem cells: A review of challenges and prospects J Integr Agric 14 222 233 10.1016/S2095-3119(14)60881-9

상세보기
Lee DY Lee SY Jung JW Kim JH Oh DH Kim HW Kang JH Choi JS Kim GD Joo ST Hur SJ 2022 Review of technology and materials for the development of cultured meat Crit Rev Food Sci Nutr 25 1 25 10.1080/10408398.2022.2063249 35466822

상세보기
Liu HS Jan MS Chou CK Chen PH Ke NJ 1999 Is green fluorescent protein toxic to the living cells? Biochem Biophys Res Commun 260 712 717 10.1006/bbrc.1999.0954 10403831

상세보기
Matsuyoshi Y Akahoshi M Nakamura M Tatsumi R Mizunoya W 2019 Isolation and purification of satellite cells from young rats by Percoll density gradient centrifugation In Myogenesis: Methods in molecular biology Rønning SB Humana Press New York, NY, USA 81 93 10.1007/978-1-4939-8897-6_6
Mauro A 1961 Satellite cell of skeletal muscle fibers J Biophys Biochem Cytol 9 493 495 10.1083/jcb.9.2.493 13768451

상세보기
Park YG Moon JH Kim J 2006 A comparative study of magnetic-activated cell sorting, cytotoxicity and preplating for the purification of human myoblasts Yonsei Med J 47 179 183 10.3349/ymj.2006.47.2.179 16642545

상세보기
Plouffe BD Murthy SK Lewis LH 2015 Fundamentals and application of magnetic particles in cell isolation and enrichment: A review Rep Prog Phys 78 016601 10.1088/0034-4885/78/1/016601 25471081

상세보기
Post MJ 2012 Cultured meat from stem cells: Challenges and prospects Meat Sci 92 297 301 10.1016/j.meatsci.2012.04.008 22543115

상세보기
Rouger K Fornasari B Armengol V Jouvion G Leroux I Dubreil L Feron M Guevel L Cherel Y 2007 Progenitor cell isolation from muscle-derived cells based on adhesion properties J Histochem Cytochem 55 607 618 10.1369/jhc.6A6954.2007 17312016

상세보기
Seale P Sabourin LA Girgis-Gabardo A Mansouri A Gruss P Rudnicki MA 2000 Pax7 is required for the specification of myogenic satellite cells Cell 102 777 786 10.1016/S0092-8674(00)00066-0 11030621

상세보기
Whitton C Bogueva D Marinova D Phillips CJC 2021 Are we approaching peak meat consumption? Analysis of meat consumption from 2000 to 2019 in 35 countries and its relationship to gross domestic product Animals 11 3466 10.3390/ani11123466 34944243

상세보기
Yoshioka K Kitajima Y Okazaki N Chiba K Yonekura A Ono Y 2020 A modified pre-plating method for high-yield and high-purity muscle stem cell isolation from human/mouse skeletal muscle tissues Front Cell Dev Biol 8 793 10.3389/fcell.2020.00793 32903486

상세보기
Zammit PS Beauchamp JR 2001 The skeletal muscle satellite cell: Stem cell or son of stem cell? Differentiation 68 193 204 10.1046/j.1432-0436.2001.680407.x 11776472

상세보기
Zammit PS Relaix F Nagata Y Ruiz AP Collins CA Partridge TA Beauchamp JR 2006 Pax7 and myogenic progression in skeletal muscle satellite cells J Cell Sci 119 1824 1832 10.1242/jcs.02908 16608873

상세보기

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