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



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