Virus Cultivation

Primary, Human Diploid, and Continuous Culture Cell Lines for Virus Isolation and Proliferation and Virus-based Vaccines

The advent of cell culture techniques has fundamentally changed virus isolation and proliferation in the lab setting. Cell-based production systems offer a convenient and cost-effective approach for the isolation, detection, and identification of viruses. Greater process control contributes to a more reliable and well-characterized product, with faster and shorter production cycles than that of animal- and egg-based production systems.

Cell-based production systems for virus culture and vaccine production are important for:

• Virus detection/identification: Cell cultures provide a suitable environment for detection and identification of many human viral pathogens, affording important microscopic examinations for evidence of viral proliferation. Accurate identification of virus is important to ensure timely and appropriate treatments, and can facilitate the detection of mixed viral infections.
• Host-pathogen interaction research: Innovations in cell biology have allowed deeper and more complex insights into host-pathogen interactions for the study of pathogenesis. In vitro cell culture systems can facilitate experimental access for investigation of the mode and etiological factors of viral infection.
• Viral structure and replication: Genetic material and replication methods vary considerably among different types of viruses. Cell culture systems can facilitate virus growth and elucidate development and interactions with host cells at every stage of replication.
• Vaccine production: Cell-based vaccine production systems offer a flexible and cost-effective approach for meeting vaccine output needs. Manufacturers can supply vaccines more quickly and in greater quantities to alleviate vaccine supply shortages during outbreaks when traditional egg-based production systems may fall short. Virus-based vaccines produced in mammalian cells may also offer better protection against viral infections, as they more closely replicate viruses in circulation than vaccines produced in chicken eggs.

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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