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Paper II — 2015 September (Supplementary) (2010 Scheme) — Q6

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Paper II
2015 September (Supplementary) (2010 Scheme) · 40 marks · 120 min

Question

Cell cultures for growing viruses

Q62 marksShort Notes

Answer

Cell cultures are one of the three classical methods for laboratory propagation/growth of viruses (alongside embryonated eggs and laboratory animal inoculation), and remain the mainstay of diagnostic virology and vaccine production.

Types of cell culture:

  1. Primary cell culture — cells freshly derived directly from animal/human tissue (e.g., primary monkey kidney cells, chick embryo fibroblasts); retains normal diploid karyotype; capable of only limited passage/subculture before the cells die out; historically important for isolating many viruses (e.g., poliovirus) and still used for certain vaccine production.
  2. Diploid cell strain (semi-continuous cell line) — a cell population (typically of human fibroblast origin, e.g., WI-38, MRC-5) that retains a normal diploid karyotype but can be subcultured for a limited, though extended, number of passages (~50–100 population doublings) before senescence; used for growing certain vaccines (e.g., some rabies, varicella vaccines) where a normal, non-transformed cell substrate is preferred for safety reasons.
  3. Continuous (established) cell line — derived from malignant tissue or transformed/immortalized cells, capable of indefinite serial passage; typically aneuploid; examples include HeLa cells (cervical carcinoma-derived) and Vero cells (monkey kidney-derived, very widely used for vaccine production and diagnostic viral culture).

Detection of viral growth in cell culture:

  • Cytopathic effect (CPE) — visible morphological changes in infected cells (rounding, syncytium formation, inclusion bodies, cell lysis), examined under light microscopy.
  • Haemadsorption — for viruses (e.g., influenza, parainfluenza) that express haemagglutinin on the infected cell surface but produce minimal visible CPE, detected by the ability of added red blood cells to adhere to the infected monolayer.
  • Interference — detecting a non-cytopathic virus by its ability to interfere with (block) the CPE of a second, known cytopathic virus subsequently added to the same culture.
  • Immunofluorescence/molecular methods — direct detection of viral antigen or nucleic acid within infected cells.

Applications: primary diagnostic virus isolation (though increasingly supplemented/replaced by faster PCR-based methods in routine practice), viral vaccine production, antiviral drug susceptibility testing, and basic virology research.

  1. 1General Properties of Viruses and Virus Host Interaction

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