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Animal Cell Culture Vessels

    In this animal cell culture vessels post we have briefly explained about common materials and alternative cell culture vessels.

    The majority of in vitro cultivated vertebrate cells form monolayers on artificial culture vessels. As a result, the substrate must be properly charged to allow cell adherence, or at the very least to allow cell-derived attachment factors to adhere, allowing cell adherence and spreading.

    Although hematopoietic cell lines, rodent ascites tumours, and a few other selected cell lines, such as human small-cell lung cancer, can grow in suspension and become independent of the surface charge on the substrate, many transformed cell lines can be made to grow in suspension and become independent of the surface charge on the culture vessels.

    Most normal cells, on the other hand, need to spread out on culture vessels to proliferate, and insufficient spreading owing to poor adhesion or overcrowding will prevent this. Anchorage dependent cells are those that require attachment for growth; transformed cells, on the other hand, frequently become anchorage independent and can grow in suspension when stirred or retained in suspension with semisolid media such as agar.

    Common Materials

    Disposable plastic

    Simple, reproducible culture vessels include single-use sterile polystyrene flasks, Petri dishes, and multiwell plates. They usually have good optical quality and a flat growing surface, resulting in monolayer cultures that are evenly dispersed and repeatable. 

    Because polystyrene is hydrophobic and does not provide a suitable surface for cell attachment when it is made, tissue culture plastics are charged, wettable surfaces are created by corona discharge, γ-irradiation, or chemical treatment.

    Glass

    Because of its optical qualities and surface charge, this was the initial culture vessels, but plastic has now supplanted it in most laboratories because to its higher uniformity and superior optical qualities. 

    Glass is no longer widely utilised as culture vessels, despite the fact that it is inexpensive, easy to clean without losing its growth-promoting characteristics, sterilizable with dry or moist heat, and optically transparent.

    Glass becomes unsuitable for culture vessels after being treated with a high alkali (e.g., NaOH or caustic detergents) until it is neutralised by an acid wash. Because high optical grade glass is alkaline and often contains a significant amount of lead, it may be necessary to acid wash and/or coat slides and coverslips for the best results.

    Animal Cell Culture Vessels

    Animal Cell Culture Vessels

    Other plastics

    Although polystyrene is by far the most common and cheapest plastic culture vessels, cells can also be grown on polyvinylchloride (PVC), polycarbonate, polytetrafluorethylene (PTFE; Teflon), Melinex, Thermanox (TPX), poly(methyl methacrylate) (PMMA; Plexiglas, Perspex, Lucite), and a number of other plastics.

    Fibers

    Rayon, Nylon, poly-L-lactic acid (PLA), polyglycolic acid (PGA), and silk are often used for two and three-dimensional constructs in tissue engineering, particularly PLA, PGA, and silk, as they are biodegradable.

    Derivatization

    Substrates that are not naturally adhesive can be derivatized with the RGD tripeptide, usually as the pentapeptide GRGDS to allow interaction with integrins on the cell surface. EDC (1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride) and N-hydroxysulfosuccinimide (sulpho-NHS) have been used to derivatize processed silk to construct scaffolds for bone tissue engineering and this treatment is potentially applicable to a number of different culture vessels.

    Metals

    Cells may be grown on stainless steel disks or other metallic surfaces. Observation of the cells on an opaque substrate requires surface interference microscopy, unless very thin metallic films are used. In 1978 Westermark developed a method for the growth of fibroblasts and glia on palladium.

    Using electron microscopy shadowing equipment, he produced islands of palladium on agarose, which does not allow cell attachment in fluid media. The size and shape of the islands were determined by masks made by photo-etching, and the palladium was applied by shadowing under vacuum, as used in electron microscopy. Because the layer was very thin, it remained transparent.

    Further Readings

    Reference

    1. Animal Cell Culture: General Considerations (biologydiscussion.com)
    2. Animal Cell Culture Guide | ATCC
    3. Animal Cell Culture: History, Types and Applications (biotechnologynotes.com)
    4. https://biochem.oregonstate.edu/content/biochemistry-free-and-easy