By John R. W. Masters
This new version of Animal mobile tradition covers new or up to date chapters on telephone authentication, serum-free tradition, apoptosis assays, FISH, genetic amendment, scale-up, stem mobile assays, three-dimensional tradition, tissue engineering and cytotoxicity assays. distinctive protocols for a large choice of equipment give you the middle of every bankruptcy, making new technique simply obtainable. every person operating in organic and scientific learn, no matter if in academia or a advertisement association, working towards phone tradition will profit drastically from this booklet.
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Extra info for Animal Cell Culture: A Practical Approach
Pyrex) is preferred because soda-lime glass releases alkali into the medium and needs to be detoxified (by boiling in weak acid) 34 SCALING-UP OF ANIMAL CELL CULTURES before use. After repeated use glassware can become less efficient for cell attachment, but efficiency can be regained by treatment with 1 mM magnesium acetate. After several hours soaking at room temperature the acetate is poured away and the glassware is rinsed with distilled water and autoclaved. ii. Plastics Polystyrene is the most used plastic for cell culture, but polyethylene, polycarbonate, Perspex, PVC, Teflon, cellophane, and cellulose acetate are all suitable when pre-treated correctly.
The answer is to carry out frequent medium changes or use perfusion, or have a pH control system. The basis of a pH control system is an autoclavable pH probe (available from Pye Ingold, Russell). This feeds a signal to the pH controller which is converted to give a digital or analogue display of the pH. This is a pH monitor system. Control of pH requires the defining of high and low pH values beyond which the pH should not go. These two set points on the pH scale turn on a relay to activate a pump, or solenoid valve, which allows additions of acid or alkali to be made to the culture to bring it back to within the allowable units.
As continuous cell lines can be genetically unstable they should only be used continuously for approximately three months, before stock replacement. Some cell lines, such as 3T3 and other mouse lines which are immortal but not transformed, can transform spontaneously, and should not be propagated for prolonged periods. Validated and authenticated frozen stocks of all cell lines should be maintained to protect against cell line instability, and to give insurance against contamination, incubator failure, or other accidental loss (see Chapter 3).