Food and Fermentation Industries >

2025 , Vol. 51 >Issue 17: 73 - 81

DOI: https://doi.org/10.13995/j.cnki.11-1802/ts.041179

Efficient isolation and culture of preadipocytes of snakehead (Channa argus) for cell-cultivated fish production

  • ZHANG Ximin ,
  • LI Yan ,
  • GUAN Xin ,
  • DU Guocheng
Expand
  • 1(Key Laboratory of Industrial Biotechnology, Ministry of Education, Jiangnan University-School of Biotechnology, Wuxi 214122, China)
    2(Science Center for Future Foods, Jiangnan University, Wuxi 214122, China)
    3(Jiangsu Basic Research Center of Synthetic Biology, Wuxi 214122, China)

Received date: 2024-09-29

  Revised date: 2025-01-08

  Online published: 2025-09-29

Fold

Abstract

To efficiently obtain seed cells and enhance the industrial production of cell-cultivated fish, an efficient and feasible method for isolation and culture of Channa argus preadipocytes was established in this study.The effects of digestion time and amount of collagenase on cell isolation efficiency were investigated, the influence of extracellular matrix on cell proliferation were assessed.The biological properties of C.argus preadipocytes were characterized through immunofluorescence staining and adipogenic induction assays, and the in vitro proliferative capacity of cells was evaluated by continuous culture.The results indicated that the highest isolation efficiency of preadipocytes was achieved using 0.25% (volume fraction) type I collagenase for tissue digestion, with an enzyme dosage of 2 mL collagenase per gram of adipose tissue and a digestion time of 45 minutes.The initial number of cells released per gram of adipose tissue reached up to 5×106, and the number of cells harvested after adherent culture was approximately 1×105 per gram of adipose tissue.Precoating with 10 g/L porcine gelatin facilitated the proliferation of preadipocytes.Following adipogenic induction, the differentiation rate of the cells was 100%, indicating that the cultured cells possess the potential to produce cultivated fat.The isolated preadipocytes could be continuously cultured in vitro for 54 days with 6 passages, achieving a 70-fold cell expansion in cell number.In summary, this study lays the groundwork for large-scale cell expansion and the future industrial production of cell-cultivated fish.

Key words： cultivated fat; cell-cultivated fish; future foods; cell separation; Channa argus; preadipocytes; collagenase

Cite this article

ZHANG Ximin , LI Yan , GUAN Xin , DU Guocheng . Efficient isolation and culture of preadipocytes of snakehead (Channa argus) for cell-cultivated fish production[J]. Food and Fermentation Industries, 2025 , 51(17) : 73 -81 . DOI: 10.13995/j.cnki.11-1802/ts.041179

References

[1] RUBIO N, DATAR I, STACHURA D, et al.Cell-based fish:A novel approach to seafood production and an opportunity for cellular agriculture[J].Frontiers in Sustainable Food Systems, 2019, 3:43.
[2] GOSWAMI M, BELATHUR SHAMBHUGOWDA Y, SATHIYANARAYANAN A, et al.Cellular aquaculture:Prospects and challenges[J].Micromachines, 2022, 13(6):828.
[3] GOSWAMI M, YASHWANTH B S, TRUDEAU V, et al.Role and relevance of fish cell lines in advanced in vitro research[J].Molecular Biology Reports, 2022, 49(3):2393-2411.
[4] XU E B, NIU R H, LAO J H, et al.Tissue-like cultured fish fillets through a synthetic food pipeline[J].NPJ Science of Food, 2023, 7:17.
[5] YIN H W, WANG L, HUR S J, et al.Cell-cultured fish meat via scale-up expansion of Carassius auratus skeletal muscle cells using edible porous microcarriers and quality evaluation[J].Journal of Agricultural and Food Chemistry, 2024, 72(29):16475-16483.
[6] GAO Y X, KONG Q, ZHENG H W, et al.Tailored edible 3D porous scaffolds constructed by Pickering emulgel templating for cell-cultured fish meat[J].Food Hydrocolloids, 2024, 157:110453.
[7] JIMÉNEZ COLMENERO F.Relevant factors in strategies for fat reduction in meat products[J].Trends in Food Science & Technology, 2000, 11(2):56-66.
[8] TOCHER D R.Metabolism and functions of lipids and fatty acids in teleost fish[J].Reviews in Fisheries Science, 2003, 11(2):107-184.
[9] 聂国兴, 傅艳茹, 张浩, 等.乌鳢肌肉营养成分分析[J].淡水渔业, 2002, 32(2):46-47.
NIE G X, FU Y R, ZHANG H, et al.Analysis of nutritional ingredient in Ophiocephalus argus's muscle[J].Freshwater Fisheries, 2002, 32(2):46-47.
[10] 谭璐. 一文读懂2023年中国乌鳢产业全景分析:预制菜等新需求带动乌鳢需求和价格增长[EB/OL].(2023-07-05)[2024-09-29].https://www.sohu.com/a/694643688_120961824.
TAN L.A panoramic analysis of China's snakehead industry in 2023:New demand such as prepared dishes drives the growth of the demand and price of snakehead[EB/OL].(2023-07-05)[2024-09-29].https://www.sohu.com/a/694643688_120961824.
[11] ANDREASSEN R C, RØNNING S B, SOLBERG N T, et al.Production of food-grade microcarriers based on by-products from the food industry to facilitate the expansion of bovine skeletal muscle satellite cells for cultured meat production[J].Biomaterials, 2022, 286:121602.
[12] 田志华, 杨公社, 赵兴波, 等.大鼠前体脂肪细胞分化过程中波形纤维形态结构的变化和波形纤维蛋白基因表达的研究[J].遗传学报, 2003, 30(12):1113-1117.
TIAN Z H, YANG G S, ZHAO X B, et al.Studies of the gene expression and structure alteration of vimentin during differentiation of rat preadipocytes[J].Acta Genetica Sinica, 2003, 30(12):1113-1117.
[13] RITTER A, FRIEMEL A, ROTH S, et al.Subcutaneous and visceral adipose-derived mesenchymal stem cells:Commonality and diversity[J].Cells, 2019, 8(10):1288.
[14] YUEN J S K Jr, STOUT A J, KAWECKI N S, et al.Perspectives on scaling production of adipose tissue for food applications[J].Biomaterials, 2022, 280:121273.
[15] 陶凯, 刘晓燕, 梁久龙, 等.人脂肪组织来源干细胞体外培养条件的优化[J].中国美容整形外科杂志, 2007, 18(3):223-226.
TAO K, LIU X Y, LIANG J L, et al.The optimization of culture conditions for human adipose tissue-derived stem cells cultured in vitro[J].Chinese Journal of Aesthetic and Plastic Surgery, 2007, 18(3):223-226.
[16] 王雅文, 乔芳, 张美玲, 等.外源性脂肪酸对罗非鱼脂肪细胞增殖与分化的影响[J].水生生物学报, 2018, 42(3):517-524.
WANG Y W, QIAO F, ZHANG M L, et al.Effects of exogenous fatty acid on the proliferation and differentiation of Nile tilapia preadipocytes[J].Acta Hydrobiologica Sinica, 2018, 42(3):517-524.
[17] 吉红, 曹艳姿, 林亚秋, 等.草鱼前体脂肪细胞的原代培养[J].水生生物学报, 2009, 33(6):1226-1230.
JI H, CAO Y Z, LIN Y Q, et al.Primary culture of grass carp preadipocyte in vitro[J].Acta Hydrobiologica Sinica, 2009, 33(6):1226-1230.
[18] VEGUSDAL A, SUNDVOLD H, GJØEN T, et al.An in vitro method for studying the proliferation and differentiation of Atlantic salmon preadipocytes[J].Lipids, 2003, 38(3):289-296.
[19] WANG X X, HUANG M, WANG Y Z.The effect of insulin, TNFα and DHA on the proliferation, differentiation and lipolysis of preadipocytes isolated from large yellow croaker (Pseudosciaena crocea R.)[J].PLoS One, 2012, 7(10):e48069.
[20] EMONT M P, JACOBS C, ESSENE A L, et al.A single-cell atlas of human and mouse white adipose tissue[J].Nature, 2022, 603(7903):926-933.
[21] 刘茜. 草鱼LPL基因克隆及草鱼脂肪细胞提取与培养的研究[D].杨凌:西北农林科技大学, 2007.
LIU Q.The research on cloning of LPL and the extraction and culture of adipocytes in grass carp[D].Yangling:Northwest A&F University, 2007.
[22] HARRIS L J, ZHANG P, ABDOLLAHI H, et al.Availability of adipose-derived stem cells in patients undergoing vascular surgical procedures[J].Journal of Surgical Research, 2010, 163(2):e105-12.
[23] VAN HARMELEN V, SKURK T, RÖHRIG K, et al.Effect of BMI and age on adipose tissue cellularity and differentiation capacity in women[J].International Journal of Obesity, 2003, 27(8):889-895.
[24] GOSWAMI M, OVISSIPOUR R, BOMKAMP C, et al.Cell-cultivated aquatic food products:Emerging production systems for seafood[J].Journal of Biological Engineering, 2024, 18(1):43.
[25] LIN L, REGENSTEIN J M, LV S, et al.An overview of gelatin derived from aquatic animals:Properties and modification[J].Trends in Food Science & Technology, 2017, 68:102-112.
[26] WU J T, XIAO J, ZHU M Y, et al.Study of physicochemical and gelation properties of fish gelatin from different sources[J].Applied Sciences, 2023, 13(9):5337.
[27] PELHAM R J, CHANG F.Role of actin polymerization and actin cables in actin-patch movement in Schizosaccharomyces pombe[J].Nature Cell Biology, 2001, 3(3):235-244.
[28] 樊廷俊, 田梦, 赵君.细胞外基质对细胞行为调控作用的研究进展[J].生命科学, 2021, 33(7):844-852.
FAN T J, TIAN M, ZHAO J.Progress in the regulation of extracellular matrix on cell behavior[J].Chinese Bulletin of Life Sciences, 2021, 33(7):844-852.
[29] 杨红文. 猪前脂肪细胞的分离培养与鉴定及猪SMAF1,ATGL基因的组织表达规律[D].武汉:华中农业大学, 2006.
YANG H W.Primary culture and identification of porcine preadipocyte and tissue expression distribution of porcine SMAF1, ATGL gene[D].Wuhan:Huazhong Agricultural University, 2006.
[30] BHARTI D, SHIVAKUMAR S B, SUBBARAO R B, et al.Research advancements in porcine derived mesenchymal stem cells[J].Current Stem Cell Research & Therapy, 2016, 11(1):78-93.
[31] HILL A B T, BRESSAN F F, MURPHY B D, et al.Applications of mesenchymal stem cell technology in bovine species[J].Stem Cell Research & Therapy, 2019, 10(1):44.
[32] JUREK S, SANDHU M A, TRAPPE S, et al.Optimizing adipogenic transdifferentiation of bovine mesenchymal stem cells:A prominent role of ascorbic acid in FABP4 induction[J].Adipocyte, 2020, 9(1):35-50.
[33] LU T F, PEI W H, WANG K F, et al.In vitro culture and biological properties of broiler adipose-derived stem cells[J].Experimental and Therapeutic Medicine, 2018, 16(3):2399-2407.
[34] 王新霞, 汪以真.大黄鱼脂肪细胞体外培养方法:中国,CN102140439B[P].2013-05-01.
WANG X X, WANG Y Z.Methods for in vitro culture of large yellow croaker adipocytes:China, CN102140439B[P].2013-05-01.
[35] 柯轶雄, 毛孝江, 任惠娟, 等.花鲈前脂肪细胞系的建立及油酸诱导分化[J].水生生物学报, 2024, 48(7):1102-1110.
KE Y X, MAO X J, REN H J, et al.Establishment of a preadipocyte cell line of spotted sea bass(Lateolabrax maculatus) and oleic acid-induced differentiation[J].Acta Hydrobiologica Sinica, 2024, 48(7):1102-1110.
[36] 温加静. 罗非鱼前脂肪细胞传代培养模型的建立及诱导分化[D].上海:华东师范大学, 2019.
WEN J J.Establishment of tilapia preadipocyte subculture model and induction of differentiation[D].Shanghai:East China Normal University, 2019.
Options
Outlines

/

〈 〉

Powered by Beijing Magtech Co. Ltd,.