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    8.Virology-Cell Culture

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    Abdus Subhan
    Viruses require living host cells to replicate and are often cultivated using appropriate cell cultures. Animal cell cultures are derived from tissue samples treated with enzymes to dissociate the cells. The cells are grown as a monolayer with a culture medium and antibiotics to prevent bacterial contamination. There are three main types of cell lines: primary cell lines which most closely represent the original tissue but have a limited lifespan; diploid cell lines which can undergo a finite number of divisions before senescing; and continuous cell lines which can be propagated indefinitely but have lost their original characteristics.

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    8.Virology-Cell Culture

    Uploaded by

    Abdus Subhan
    25 views2 pages
    Viruses require living host cells to replicate and are often cultivated using appropriate cell cultures. Animal cell cultures are derived from tissue samples treated with enzymes to dissociate the cells. The cells are grown as a monolayer with a culture medium and antibiotics to prevent bacterial contamination. There are three main types of cell lines: primary cell lines which most closely represent the original tissue but have a limited lifespan; diploid cell lines which can undergo a finite number of divisions before senescing; and continuous cell lines which can be propagated indefinitely but have lost their original characteristics.

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    Viruses require living host cells to replicate and are often cultivated using appropriate cell cultures. Animal cell cultures are derived from tissue samples treated with enzymes to dissociate the cells. The cells are grown as a monolayer with a culture medium and antibiotics to prevent bacterial contamination. There are three main types of cell lines: primary cell lines which most closely represent the original tissue but have a limited lifespan; diploid cell lines which can undergo a finite number of divisions before senescing; and continuous cell lines which can be propagated indefinitely but have lost their original characteristics.

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    © All Rights Reserved
    Download as PDF, TXT or read online from Scribd
    Download as pdf or txt
    25 views2 pages

    8.Virology-Cell Culture

    Uploaded by

    Abdus Subhan
    Viruses require living host cells to replicate and are often cultivated using appropriate cell cultures. Animal cell cultures are derived from tissue samples treated with enzymes to dissociate the cells. The cells are grown as a monolayer with a culture medium and antibiotics to prevent bacterial contamination. There are three main types of cell lines: primary cell lines which most closely represent the original tissue but have a limited lifespan; diploid cell lines which can undergo a finite number of divisions before senescing; and continuous cell lines which can be propagated indefinitely but have lost their original characteristics.

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    © All Rights Reserved
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    of 2

    CELL CULTURE

    Because viruses replicate only inside living cells, the cultivation of viruses requires
    the use of appropriate hosts. Viruses infecting prokaryotes are typically the easiest
    to grow in the laboratory. For the study of bacterial viruses, pure cultures are used
    either in liquid or on semisolid (agar) media. Most animal viruses and many plant
    viruses can be cultivated in tissue or cell cultures, and the use of such cultures has
    enormously facilitated research on these viruses. Plant viruses can be more difficult
    to work with, because their study sometimes requires use of the whole plant. This is
    a problem because plants grow much slower than bacteria, and plant viruses also
    often require a break in the thick plant cell wall in order to infect.

    Cell Culture Preparation


    Animal cell cultures are derived from cells originally taken from an organ of an
    experimental animal. Unless blood cells are used, cell cultures are usually obtained
    by aseptically removing pieces of tissue and dissociating the cells by treatment with
    an enzyme that degrades the extracellular material that holds animal cells together.
    The resulting cell suspension is spread over a flat surface, such as the bottom of a
    culture flask or a Petri dish. The thin layer of cells adhering to the glass or plastic
    dish, called a monolayer, is overlaid with a suitable culture medium and incubated
    at a suitable temperature. The culture media used for cell cultures are typically quite
    complex, containing a number of amino acids and vitamins, salts, glucose, and a
    bicarbonate buffer system. To obtain the best growth, addition of a small amount of
    blood serum is usually necessary to provide vital nutrients, and several antibiotics
    are added to prevent bacterial contamination.

     Primary Cell Line


    Primary cells most closely represent the tissue of origin. They are taken directly from
    the tissue and processed to establish them under optimized culture conditions.
    Because they are derived from tissue and not modified, they are more similar to the
    in vivo state and exhibit normal physiology. For this reason, they provide excellent
    model systems for studying the normal physiology and biochemistry of cells (e.g.,
    metabolic studies, aging, signaling studies), and the effects of drugs and toxic
    compounds on the cells. Keep in mind that primary cells have a limited lifespan and
    will stop dividing (or senesce) after a certain number of cell divisions and can be
    more difficult to culture and maintain than a continuous cell line. The variability
    induced in primary cells acquired from donors and during subculture practices is a
    major challenge faced by the researchers who study cell signaling pathways.
    Researchers prefer to screen the cells for sensitivity to common stimuli before
    embarking on signaling studies. Pre-screened primary cells save valuable resources
    as they are stimulated for activation of major signaling pathways. The most popular
    types of primary cells used in research are epithelial cells, fibroblasts, keratinocytes,
    melanocytes, endothelial cells, muscle cells, hematopoietic and mesenchymal stem
    cells.

     Diploid Cell Line


    Diploid cultures have a finite lifespan. They usually can undergo a maximum of 20-
    80 PDLs (Population Doubling Level) before they senesce. Normal human cells,
    such as human skin fibroblasts, are one example of diploid cells. Most widely used
    cells are those established from human embryonic lung such as WI-38.

     Continuous Cell Line


    Continuous immortalized cell lines are comprised of a single cell type that can be
    serially propagated in culture either for a limited number of cell divisions
    (approximately thirty) or otherwise indefinitely. Cell lines of a finite life are usually
    diploid and maintain some degree of differentiation. The fact that such cell lines
    senesce after approximately thirty cycles of division means it is essential to establish
    a system of Master and Working banks in order to maintain such lines for long
    periods. Continuous immortalized cell lines that can be propagated indefinitely
    generally have this ability because they have been transformed into tumor cells.
    Tumor cell lines are often derived from actual clinical tumors, but transformation
    may also be induced using viral oncogenes or by chemical treatments. Transformed
    cell lines present the advantage of almost limitless availability, but the disadvantage
    of having retained very little of the original in vivo characteristics. Most widely used
    continuous cells are Hep-2 and HeLa cells derived from human carcinomas.

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