Novel Eel Skin Fibroblast Cell Line: Bridging Adherent and Suspension Growth for Aquatic Applications Including Virus Susceptibility

Overview

Journal Biology (Basel)

Publisher MDPI

Specialty Biology

Date 2025 Jan 8

PMID 39765736

Authors

Zaiyu Zheng

Bin Chen

Xiaodong Liu

Rui Guo

Hongshu Chi

Xiuxia Chen

Ying Pan

Hui Gong

Affiliations

Soon will be listed here.

Abstract

Suspension growth can greatly increase the cell density and yield of cell metabolites. To meet the requirements of aquatic industries, a culture model derived from skin was developed using the explant outgrowth and enzyme-digesting passaging methods. These cells were kept in vitro continuously for over 12 months and subcultured 68 times. This heteroploid cell line, designated as ES, can naturally adapt to adherent and suspension growth reversibly under certain temperatures, serum percentages, and inoculum densities, without the need for any microcarriers or special medium additives. The ES cells can continue being highly productive under a temperature range of 15-37 °C and a serum percentage ranging from 3 to 15%. An inoculum density higher than 5 × 10 cells·mL is necessary for the ES cells to turn into suspension efficiently. The green fluorescent protein (GFP) reporter gene was successfully expressed in the ES cells. The ES cells demonstrated susceptibility to Anguillid herpesvirus (AngHV) and red-spotted grouper nervous necrosis virus (RGNNV). ES is the first natural suspension growth model of aquatic origin; it does not require the processes of suspension domestication and carrier dissolution, making it a promising and cost-effective model for vaccine production, bio-pharmaceutical manufacturing, and cellular agriculture.

References

Chang P, Pan Y, Wu C, Kuo S, Chung H . Isolation and molecular characterization of herpesvirus from cultured European eels Anguilla anguilla in Taiwan. Dis Aquat Organ. 2002; 50(2):111-8. DOI: 10.3354/dao050111. View

Grieger J, Soltys S, Samulski R . Production of Recombinant Adeno-associated Virus Vectors Using Suspension HEK293 Cells and Continuous Harvest of Vector From the Culture Media for GMP FIX and FLT1 Clinical Vector. Mol Ther. 2015; 24(2):287-297. PMC: 4817810. DOI: 10.1038/mt.2015.187. View

Lee N, Shin J, Park J, Lee G, Cho S, Cho B . Targeted Gene Deletion Using DNA-Free RNA-Guided Cas9 Nuclease Accelerates Adaptation of CHO Cells to Suspension Culture. ACS Synth Biol. 2016; 5(11):1211-1219. DOI: 10.1021/acssynbio.5b00249. View

Tihanyi B, Nyitray L . Recent advances in CHO cell line development for recombinant protein production. Drug Discov Today Technol. 2021; 38:25-34. DOI: 10.1016/j.ddtec.2021.02.003. View

Chu C, Lugovtsev V, Golding H, Betenbaugh M, Shiloach J . Conversion of MDCK cell line to suspension culture by transfecting with human siat7e gene and its application for influenza virus production. Proc Natl Acad Sci U S A. 2009; 106(35):14802-7. PMC: 2728112. DOI: 10.1073/pnas.0905912106. View

van Nieuwstadt A, Dijkstra S, Haenen O . Persistence of herpesvirus of eel Herpesvirus anguillae in farmed European eel Anguilla anguilla. Dis Aquat Organ. 2001; 45(2):103-7. DOI: 10.3354/dao045103. View

Ryu N, Lee S, Park H . Spheroid Culture System Methods and Applications for Mesenchymal Stem Cells. Cells. 2019; 8(12). PMC: 6953111. DOI: 10.3390/cells8121620. View

Merten O . Introduction to animal cell culture technology-past, present and future. Cytotechnology. 2008; 50(1-3):1-7. PMC: 3476009. DOI: 10.1007/s10616-006-9009-4. View

Zhang J, Qiu Z, Wang S, Liu Z, Qiao Z, Wang J . Suspended cell lines for inactivated virus vaccine production. Expert Rev Vaccines. 2023; 22(1):468-480. DOI: 10.1080/14760584.2023.2214219. View

10.

Arai T . Do we protect freshwater eels or do we drive them to extinction?. Springerplus. 2014; 3:534. PMC: 4174548. DOI: 10.1186/2193-1801-3-534. View

11.

Kumar M, Singh V, Mishra A, Kushwaha B, Kumar R, Lal K . Fish cell line: depositories, web resources and future applications. Cytotechnology. 2024; 76(1):1-25. PMC: 10828409. DOI: 10.1007/s10616-023-00601-2. View

12.

Taylor M, Weegman B, Baicu S, Giwa S . New Approaches to Cryopreservation of Cells, Tissues, and Organs. Transfus Med Hemother. 2019; 46(3):197-215. PMC: 6558330. DOI: 10.1159/000499453. View

13.

Ikeda D, Otsuka Y, Kan-No N . Development of a novel Japanese eel myoblast cell line for application in cultured meat production. Biochem Biophys Res Commun. 2024; 734:150784. DOI: 10.1016/j.bbrc.2024.150784. View

14.

Gregersen J, Schmitt H, Trusheim H, Broker M . Safety of MDCK cell culture-based influenza vaccines. Future Microbiol. 2011; 6(2):143-52. DOI: 10.2217/fmb.10.161. View

15.

Zheng Z, Yang J, Ge J, Chi H, Chen B, Fang Q . Development and characterization of a continuous cell line (EL) from the liver of European eel Anguilla anguilla. Cell Biol Int. 2019; 44(3):808-820. PMC: 7028054. DOI: 10.1002/cbin.11276. View

16.

Frankowski J, Bastrop R . Identification of Anguilla anguilla (L.) and Anguilla rostrata (Le Sueur) and their hybrids based on a diagnostic single nucleotide polymorphism in nuclear 18S rDNA. Mol Ecol Resour. 2011; 10(1):173-6. DOI: 10.1111/j.1755-0998.2009.02698.x. View

17.

Bandin I, Souto S . Betanodavirus and VER Disease: A 30-year Research Review. Pathogens. 2020; 9(2). PMC: 7168202. DOI: 10.3390/pathogens9020106. View

18.

Chen B, Zheng Z, Yang J, Chi H, Huang H, Gong H . Development and characterization of a new cell line derived from European eel kidney. Biol Open. 2018; 8(1). PMC: 6361207. DOI: 10.1242/bio.037507. View

19.

Chi S, Shieh J, Lin S . Genetic and antigenic analysis of betanodaviruses isolated from aquatic organisms in Taiwan. Dis Aquat Organ. 2003; 55(3):221-8. DOI: 10.3354/dao055221. View

20.

Vo N, Katzenback B, Kellendonk C, Duong T, Curtis T, Dixon B . Characterization of the continuous skin fibroblastoid cell line, WE-skin11f, from walleye (Sander vitreus). J Fish Dis. 2019; 42(11):1587-1599. DOI: 10.1111/jfd.13079. View