Cell Lines for Virus Culture and Vaccine Production

Primary, human diploid, and continuous culture cell lines for the propagation of viruses and virus-based vaccines

Viruses are obligate intracellular parasites that require living cells to multiply. Early efforts to expand viruses used both animals and embryonated eggs for propagation and isolation, but the advent of cell culture techniques has fundamentally changed virus propagation in the lab setting. Cell culture systems are a convenient and cost-effective for the isolation, detection, and identification of viruses.

Viruses cultivated in cells derived from hosts are important for:

  • Virus detection/identification: Cell culture is used as a platform for detection and diagnosis of infectious pathogens, including viruses. Accurate identification of viruses is important to ensure timely treatment of infected patients with appropriate antiviral drugs. Cell culture is considered the gold standard for virus isolation and detection.
  • Host-pathogen interaction research: The ability to manipulate and exploit cellular machinery contributes to the success of viral infections. In vitro cell culture systems facilitate experimental access for investigating the mode and etiological factors of viral infection.
  • Viral structure and replication: Genomic material and the method by which it is replicated varies considerably among different types of viruses. Viruses interact with the host cell at every stage of replication.  Propagation of viruses in cell culture allows us to study their growth and development.
  • Vaccine production: Cell culture systems are cost-effective and robust approaches for vaccine production. Cell culture-based vaccine production systems can help to alleviate vaccine supply shortages during pandemic outbreaks when traditional egg-production systems may fall short, and have become primary tools for the production of seasonal vaccines.  

Cell culture systems used for virus propagation may employ primary cells, semi-continuous cell lines, and continuous cell lines:

 

Primary cell lines Human diploid cells
(Semi-continuous cell lines)
Continuous cell lines
  • Derived from animal tissues. May be passaged once or twice to generate secondary cultures, but grow only for limited time. 
  • Secondary cultures (subculture of the primary culture) are similar to primary cultures in both morphology and viral susceptibility  
  • Used both for virus isolation (clinical samples) and vaccine production      
  • Derived from fetal tissues, and can be subcultured for ~50 passages 
  • Similar to primary cultures in viral susceptibility  
  • Used for vaccine production and isolation of fastidious viruses            
  • Cancer or other immortalized cells, which multiply rapidly and may be cultured indefinitely  
  • May become heteroploid on serial passage  
  • Narrower range of viral susceptibility than primary or diploid cultures  
  • Easy to adapt viruses for propagation  
  • Used for production of serologic antigens, and a source of cells for neutralizing antibody assays  

 

Cell Line Origin Infectious Agents
A549   human lung carcinoma Adenovirus,1 HSV,2 influenza,3 measles,4 mumps,4 parainfluenza,5 poliovirus,6 respiratory syncytial virus (RSV),6 rotavirus,7 Varicella zoster virus (VZV),8 metapneumovirus (MPV)9
BHK 21 (clone 13) Syrian hamster kidney Human adenovirus D,10 reovirus 3,11 vesicular stomatitis virus (Indiana strain),12 Dengue,13 influenza,13 rabies,13 foot and mouth,14 rubella,15
CV-1   African green monkey kidney fibroblast RSV,16, measles,16, HSV,17, VZV,18
HeLa human cervix adenocarcinoma   Poliovirus type I,19 adenovirus type 3,20 CMV,21 echovirus,22 HSV23 poliovirus,24 rhinovirus,25 vesicular stomatitis (Indiana Strain) virus,26 VZV27
LLCMK2 Rhesus monkey kidney Poliovirus type 1,17 enterovirus,28 rhinovirus,29 poxvirus groups30
McCoy Mouse fibroblast HSV32
MDCK   Madin-Darby canine kidney Influenza A,31 influenza B, some types of adenovirus,33 reoviruses34
MRC-5 human fetal lung CMV,35 HSV,36 adenovirus,37 influenza,38 mumps,39 echovirus,40 poliovirus,41 rhinovirus,42 RSV,43 VZV,27
NCI-H292   Human lung, mucoepidermoid carcinoma Vaccinia virus,44 HSV,45 adenovirus,46 measles virus,47 reoviruses,48 BK polyomavirus,49 RSV,50 some strains of influenza A,51 most enteroviruses,52 and rhinoviruses53
Vero African green monkey kidney Coxsackie B,54 HSV,55 measles,56 mumps,57 poliovirus type 3,58 rotavirus,59 rubella,60
Vero76 African green monkey kidney Coxsackie B,61 HSV,62 West Nile virus,63
Wi 38 Human fetal lung Adenovirus,64 CMV,65 echovirus,66 HSV,67 mumps,68 influenza,69 rhinovirus,70 RSV,71 VZV,72

 

References

  1. Hodzic, Jasmina, et al. "Functional screening identifies human miRNAs that modulate adenovirus propagation in prostate cancer cells." Human gene therapy 28.9 (2017): 766-780.
  2. Wang, YuTao, et al. "Inhibition of influenza virus via a sesquiterpene fraction isolated from Laggera pterodonta by targeting the NF-κB and p38 pathways." BMC complementary and alternative medicine 17.1 (2017): 25.
  3. Holzberg, Magdalena, et al. "Vemurafenib limits influenza a virus propagation by targeting multiple signaling pathways." Frontiers in microbiology 8 (2017): 2426.
  4. Franz, Stephanie, et al. "Mumps Virus SH protein inhibits NF-κB activation by interacting with TNFR1, IL-1R1, and TLR3 complexes." Journal of virology (2017): JVI-01037.
  5. Dirr, Larissa, et al. "The impact of the butterfly effect on human parainfluenza virus haemagglutinin-neuraminidase inhibitor design." Scientific reports 7.1 (2017): 4507.
  6. McCaskill, Jana L., et al. "Broad-spectrum inhibition of respiratory virus infection by microRNA mimics targeting p38 MAPK signaling." Molecular Therapy-Nucleic Acids 7 (2017): 256-266.
  7. Mackowiak, Martin, et al. "Distribution of Escherichia coli, coliphages and enteric viruses in water, epilithic biofilms and sediments of an urban river in Germany." Science of The Total Environment 626 (2018): 650-659.
  8. Aiba, Nobuyasu, et al. "Interaction of Immunoglobulin with Cytomegalovirus-Infected Cells." Viral immunology 30.7 (2017): 500-507.
  9. Li, Youxian, et al. "Characterization of signaling pathways regulating the expression of pro-inflammatory long form thymic stromal lymphopoietin upon human metapneumovirus infection." Scientific reports 8.1 (2018): 883.
  10. Okeke, Malachy, et al. "Hazard Characterization of Modified Vaccinia Virus Ankara Vector: What Are the Knowledge Gaps?." Viruses 9.11 (2017): 318.
  11. Pudupakam, Raghavendra Sumanth, et al. "Genetic characterization of the non-structural protein-3 gene of bluetongue virus serotype-2 isolate from India." Veterinary world 10.3 (2017): 348.
  12. Kleinlützum, Dina, et al. "Enhancing the oncolytic activity of CD133-targeted measles virus: receptor extension or chimerism with vesicular stomatitis virus are most effective." Frontiers in oncology 7 (2017): 127.
  13. Nikolay, Alexander, et al. "Propagation of Brazilian Zika virus strains in static and suspension cultures using Vero and BHK cells." Vaccine 36.22 (2018): 3140-3145.
  14. Kamal, Samia Ahmed, and Randa Abd El-Rahman Hassan. "Advanced Virological and Clinicopathological Studies On Cattle Suffering From Foot and Mouth Disease Virus." Journal of Immunization 1.1 (2017): 33.
  15. Prescott, Joseph, Heinz Feldmann, and David Safronetz. "Amending Koch's postulates for viral disease: When “growth in pure culture” leads to a loss of virulence." Antiviral research 137 (2017): 1-5.
  16. Hampton, Cheri M., et al. "Correlated fluorescence microscopy and cryo-electron tomography of virus-infected or transfected mammalian cells." nature protocols 12.1 (2017): 150.
  17. Dotti, Silvia, et al. "Transformation and tumorigenicity testing of simian cell lines and evaluation of poliovirus replication." PloS one 12.1 (2017): e0169391.
  18. Yang, Kui, Xiaoqun Dang, and Joel D. Baines. "A domain of Herpes simplex virus pUL33 required to release monomeric viral genomes from cleaved concatameric DNA." Journal of virology (2017): JVI-00854.
  19. Gromeier, Matthias, and Smita K. Nair. "Recombinant poliovirus for cancer immunotherapy." Annual review of medicine 69 (2018): 289-299.
  20. Weigert, Melanie, et al. "RIPK3 promotes adenovirus type 5 activity." Cell death & disease 8.12 (2017): 3206.
  21. Khadivjam, Bita, et al. "The ATP-dependent RNA helicase DDX3X modulates herpes simplex virus type 1 gene expression." Journal of virology (2017): JVI-02411.
  22. Aguilera, Elizabeth R., et al. "Plaques formed by mutagenized viral populations have elevated coinfection frequencies." MBio 8.2 (2017): e02020-16.
  23. Ibáñez, Francisco J., et al. "Pharmacological Induction of Heme Oxygenase-1 Impairs Nuclear Accumulation of Herpes Simplex Virus Capsids upon Infection." Frontiers in microbiology 8 (2017): 2108.
  24. Acevedo, Ashley, et al. "Genetic recombination of poliovirus facilitates subversion of host barriers to infection." bioRxiv (2018): 273060.
  25. Ganjian, Haleh, et al. "ICAM-1 binding rhinoviruses enter HeLa cells via multiple pathways and travel to distinct intracellular compartments for uncoating." Viruses 9.4 (2017): 68.
  26. Pan, Wei, et al. "EIF3i affects vesicular stomatitis virus growth by interacting with matrix protein." Veterinary microbiology 212 (2017): 59-66.
  27. Sadaoka, Tomohiko, et al. "Human embryonic stem cell-derived neurons are highly permissive for varicella-zoster virus lytic infection." Journal of virology 92.1 (2018): e01108-17.
  28. Betancourt, Walter Q., Sherif Abd-Elmaksoud, and Charles P. Gerba. "Efficiency of Reovirus Concentration from Water with Positively Charged Filters." Food and environmental virology (2018): 1-3.
  29. Sato, Ko, et al. "Efficient isolation of human metapneumovirus using MNT‐1, a human malignant melanoma cell line with early and distinct cytopathic effects." Microbiology and immunology 61.11 (2017): 497-506.
  30. Parker, Scott, et al. "Evaluation of Taterapox Virus in Small Animals." Viruses 9.8 (2017): 203.
  31. Petyaev, Ivan M., et al. "Resveratrol Inhibits Propagation of Chlamydia trachomatis in McCoy Cells." BioMed research international 2017 (2017).
  32. Danaher, Robert J., et al. "HSV-1 clinical isolates with unique in vivo and in vitro phenotypes and insight into genomic differences." Journal of neurovirology 23.2 (2017): 171-185.
  33. Glatthaar‐Saalmüller, Bernadette, Kerstin H. Mair, and Armin Saalmüller. "Antiviral activity of aspirin against RNA viruses of the respiratory tract—an in vitro study." Influenza and other respiratory viruses 11.1 (2017): 85-92.
  34. Shah Mahmud, Raihan, et al. "Ribonuclease from bacillus acts as an antiviral agent against negative-and positive-sense single stranded human respiratory RNA viruses." BioMed research international 2017 (2017).
  35. Hobom, Urs, et al. "Fast screening procedures for random transposon libraries of cloned herpesvirus genomes: mutational analysis of human cytomegalovirus envelope glycoprotein genes." Journal of virology 74.17 (2000): 7720-7729.
  36. Didcock, L., et al. "Sendai virus and simian virus 5 block activation of interferon-responsive genes: importance for virus pathogenesis." Journal of virology 73.4 (1999): 3125-3133.
  37. Gugala, Z., et al. "Osteoinduction by ex vivo adenovirus-mediated BMP2 delivery is independent of cell type." Gene therapy 10.16 (2003): 1289.
  38. Cheng, Pei‐Win, et al. "Antiviral effects of saikosaponins on human coronavirus 229E in vitro." Clinical and experimental pharmacology and physiology 33.7 (2006): 612-616.
  39. Wilson, Michael R., et al. "A novel cause of chronic viral meningoencephalitis: C ache Valley virus." Annals of neurology 82.1 (2017): 105-114.
  40. Atieh, Thérèse, et al. "Haiku: New paradigm for the reverse genetics of emerging RNA viruses." PloS one 13.2 (2018): e0193069.
  41. Lewandowska, Dagmara W., et al. "Optimization and validation of sample preparation for metagenomic sequencing of viruses in clinical samples." Microbiome 5.1 (2017): 94.
  42. Jenny, Robert A., et al. "Productive Infection of Human Embryonic Stem Cell‐Derived NKX2. 1+ Respiratory Progenitors With Human Rhinovirus." Stem cells translational medicine 4.6 (2015): 603-614.
  43. Jerome, Keith R., ed. Lennette's laboratory diagnosis of viral infections. CRC Press, 2016.
  44. Pereira, Leonel. Therapeutic and Nutritional Uses of Algae. CRC Press, 2018.
  45. Mu, Jinmin, et al. "A Novel High-Mannose Specific Lectin from the Green Alga Halimeda renschii Exhibits a Potent Anti-Influenza Virus Activity through High-Affinity Binding to the Viral Hemagglutinin." Marine drugs 15.8 (2017): 255.
  46. Costello, Michael J., Nancy T. Smernoff, and Margaret Yungbluth. "Laboratory diagnosis of viral respiratory tract infections." Laboratory Medicine 24.3 (1993): 150-157.
  47. Currier, Michael G., et al. "EGFR interacts with the fusion protein of respiratory syncytial virus strain 2-20 and mediates infection and mucin expression." PLoS pathogens 12.5 (2016): e1005622.
  48. Tan, Bing, et al. "Isolation and characterization of adenoviruses infecting endangered golden snub-nosed monkeys (Rhinopithecus roxellana)." Virology journal 13.1 (2016): 190.
  49. Yang, Tao, et al. "Resveratrol, sirtuins, and viruses." Reviews in medical virology 25.6 (2015): 431-445.
  50. Gagliardi, Talita Bianca, et al. "Syncytia Induction by Clinical Isolates of Human Respiratory Syncytial Virus A." Intervirology 60.1-2 (2017): 56-60.
  51. Amatore, Donatella, et al. "Influenza virus replication in lung epithelial cells depends on redox‐sensitive pathways activated by NOX4‐derived ROS." Cellular microbiology 17.1 (2015): 131-145.
  52. Castells, Empar, Velma G. George, and J. C. Hierholzer. "NCI-H292 as an alternative cell line for the isolation and propagation of the human paramyxoviruses." Archives of virology 115.3-4 (1990): 277-288.
  53. Bianco, Andrea, et al. "Th2 cytokines exert a dominant influence on epithelial cell expression of the major group human rhinovirus receptor, ICAM-1." European Respiratory Journal 12.3 (1998): 619-626.
  54. Mall, Anwar Suleman, et al. "Mucus and Mucins: do they have a role in the inhibition of the human immunodeficiency virus?." Virology journal 14.1 (2017): 192.
  55. Elion, Gertrude B., et al. "Selectivity of action of an antiherpetic agent, 9-(2-hydroxyethoxymethyl) guanine." Proceedings of the National Academy of Sciences 74.12 (1977): 5716-5720.
  56. Rima, B. K., W. B. Davidson, and S. J. Martin. "The role of defective interfering particles in persistent infection of Vero cells by measles virus." Journal of General Virology 35.1 (1977): 89-97.
  57. Gouma, Sigrid, et al. "Mumps-specific cross-neutralization by MMR vaccine-induced antibodies predicts protection against mumps virus infection." Vaccine 34.35 (2016): 4166-4171.
  58. Sanders, Barbara P., et al. "Production of high titer attenuated poliovirus strains on the serum-free PER. C6® cell culture platform for the generation of safe and affordable next generation IPV." Vaccine 33.48 (2015): 6611-6616.
  59. Kang, Joo Yeon, et al. "Antiviral effects of Lactobacillus ruminis SPM0211 and Bifidobacterium longum SPM1205 and SPM1206 on rotavirus-infected Caco-2 cells and a neonatal mouse model." Journal of Microbiology 53.11 (2015): 796-803.
  60. Prasad, Vidya Mangala, Thomas Klose, and Michael G. Rossmann. "Assembly, maturation and three-dimensional helical structure of the teratogenic rubella virus." PLoS pathogens 13.6 (2017): e1006377.
  61. Venturi, Giulietta, et al. "An autochthonous case of Zika due to possible sexual transmission, Florence, Italy, 2014." Eurosurveillance 21.8 (2016).
  62. Criscuolo, Elena, et al. "Synergy evaluation of anti-Herpes Simplex Virus type 1 and 2 compounds acting on different steps of virus life cycle." Antiviral research 151 (2018): 71-77.
  63. Richard, Audrey Stéphanie, et al. "AXL-dependent infection of human fetal endothelial cells distinguishes Zika virus from other pathogenic flaviviruses." Proceedings of the National Academy of Sciences (2017): 201620558.
  64. Kim, Young Yeon, et al. "Cooperation between p21 and Akt is required for p53‐dependent cellular senescence." Aging cell 16.5 (2017): 1094-1103.
  65. Plotkin, Stanley. "The history of vaccination against cytomegalovirus." Medical microbiology and immunology 204.3 (2015): 247-254.
  66. Mintz, Lawrence, and W. Lawrence Drew. "Relation of culture site to the recovery of nonpolio enteroviruses." American journal of clinical pathology 74.3 (1980): 324-326.
  67. Baker, David A., Bruce Kleger, and Stanley A. Plotkin. "A Comparison of Herpes Simplex Virus (HSV) Typing Using Direct Fluorescent Antibody (DFA) and Chick Embryo Plaque (CEP) Assays." Laboratory Medicine 10.9 (2016): 559-561.
  68. Perelygina, Ludmila, et al. "Rubella persistence in epidermal keratinocytes and granuloma M2 macrophages in patients with primary immunodeficiencies." Journal of Allergy and Clinical Immunology 138.5 (2016): 1436-1439.
  69. Sousa, Filipa Henderson, et al. "Cathelicidins display conserved direct antiviral activity towards rhinovirus." Peptides 95 (2017): 76-83.
  70. Shariff, Sami, et al. "Human rhinovirus infection of epithelial cells modulates airway smooth muscle migration." American journal of respiratory cell and molecular biology 56.6 (2017): 796-803.
  71. Corry, Jacqueline, et al. "Preventing cleavage of the respiratory syncytial virus attachment protein in vero cells rescues the infectivity of progeny virus for primary human airway cultures." Journal of virology 90.3 (2016): 1311-1320.
  72. Zahoor, Muhammad Asif, et al. "Cell culture-based viral vaccines: current status and future prospects." Future Virology 11.7 (2016): 549-562.