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    Cell Culturing: A Beginner's Guide To Understanding The Basics of Cell Culturing

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    mieayam234
    This document provides an overview of cell culturing basics, including the two main types of cultured cells: primary cells and cell lines. Primary cells are derived directly from tissue and have a heterogeneous mix of cells, while cell lines are derived from primary cells that have been manipulated to last longer in culture. Proper aseptic technique is important to prevent bacterial and fungal contamination, including using sterile equipment and working in a laminar flow cabinet. If contamination occurs, the infected cells may be treated with antibiotics or antifungals if deemed valuable.

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    Cell Culturing: A Beginner's Guide To Understanding The Basics of Cell Culturing

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    mieayam234
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    This document provides an overview of cell culturing basics, including the two main types of cultured cells: primary cells and cell lines. Primary cells are derived directly from tissue and have a heterogeneous mix of cells, while cell lines are derived from primary cells that have been manipulated to last longer in culture. Proper aseptic technique is important to prevent bacterial and fungal contamination, including using sterile equipment and working in a laminar flow cabinet. If contamination occurs, the infected cells may be treated with antibiotics or antifungals if deemed valuable.

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    978-1-84628-740-4_4

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    This document provides an overview of cell culturing basics, including the two main types of cultured cells: primary cells and cell lines. Primary cells are derived directly from tissue and have a heterogeneous mix of cells, while cell lines are derived from primary cells that have been manipulated to last longer in culture. Proper aseptic technique is important to prevent bacterial and fungal contamination, including using sterile equipment and working in a laminar flow cabinet. If contamination occurs, the infected cells may be treated with antibiotics or antifungals if deemed valuable.

    Copyright:

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

    Cell Culturing: A Beginner's Guide To Understanding The Basics of Cell Culturing

    Uploaded by

    mieayam234
    This document provides an overview of cell culturing basics, including the two main types of cultured cells: primary cells and cell lines. Primary cells are derived directly from tissue and have a heterogeneous mix of cells, while cell lines are derived from primary cells that have been manipulated to last longer in culture. Proper aseptic technique is important to prevent bacterial and fungal contamination, including using sterile equipment and working in a laminar flow cabinet. If contamination occurs, the infected cells may be treated with antibiotics or antifungals if deemed valuable.

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    © All Rights Reserved
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    of 2

    Chapter 4

    Cell Culturing: A Beginner’s Guide


    to Understanding the Basics of
    Cell Culturing
    Khurshid Alam, Edwin T. Anthony, P.N. Vaiude, Faruquz Zaman,
    and Harshad A. Navsaria

    INTRODUCTION
    The fundamental aspect of cell culturing is to understand the
    type of cells that the investigator wishes to grow. There are many
    differences between the cell types; however, the easiest method
    of categorizing them is into primary cells and cell lines.
    Two types of cultured cell types:

    • Primary cells
    • Cell lines

    Primary Cells
    These are cells derived directly from tissue samples/biopsies
    which have heterogeneous nature of cells with variable growth
    friction. The cells are extracted directly from the tissue and
    grown directly in specified optimal cell culture Medias.

    Cell Lines
    These are cells subcultured from primary cells but now have been
    manipulated in the laboratory so that they last longer and can
    go through more cell passages with growth friction of 80% or
    more, than the original primary cells before they change their
    morphology. On the whole, they tend to be more resilient than
    primary cells. However, due to their manipulation, they may not
    mimic in-vivo cells as closely as the unmanipulated primary cells.
    Subcultured cells separated from primary cells create homoge-
    neous cell lineages. They ascertain specific properties by the
    process of cloning or physical cell separation, thus leading to
    so-called cell strains.
    32 BASIC SCIENCE TECHNIQUES IN CLINICAL PRACTICE

    FUNDAMENTALS OF CELL CULTURING


    The reason for undertaking cell culturing is that the growth of
    the cells can be mimicked as closely as possible to its growth in
    its normal host environment. By growing the cells in cell culture
    flasks or cell culture dishes, the investigator may build up the
    cell numbers and then use these cells to conduct experiments.
    These in-vitro cell experiments may be performed, and later the
    final effects on the cells may be investigated.
    The cells are delicate and prone to bacterial and fungal
    infections from the environment, so all precautions are used to
    minimize contamination. Bacterial contamination is a difficult
    problem. In this respect, mycoplasma-species contamination
    is a more difficult problem to deal with than the growth of
    other bacteria and fungi. To combat contamination, the single-
    use disposable equipment and aseptic techniques are applied
    (Figure 4.1).
    Methods to prevent cell contamination:

    • Gloves
    • Sterilized equipment
    • Laminar flow cabinet
    • Singe-se pipettes
    • Resterilization of equipment

    What Happens When an Infection Occurs?


    Usually a bacterial or fungal infection would mean that the cells
    would need to be safely discarded to prevent cross contamina-
    tion. However, when the cells are “precious,” an attempt to treat
    the contamination may be made. Free-floating contaminants in
    the media are physically removed, and the cells are washed
    numerous times with phosphate buffered saline (PBS). Loose
    lids or covers are replaced with new sterile ones. An antibiotic
    mix is added to the media, and the antibiotic medium is added
    to the cells. The cells are then cultured for 2 to 3 days. If the cells
    remain uncontaminated, then they are returned back to a normal
    media for another 2 to 3 days to confirm that no contamination
    remains.
    First-choice antibiotics/antifungal agents for cell culture:

    • Gentamicin
    • Streptomycin
    • Penicillin G
    • Amphotericin B (antifungal)

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