By John R. Masters (Editor)
This re-creation of Animal phone tradition covers new or up to date chapters on cellphone authentication, serum-free tradition, apoptosis assays, FISH, genetic amendment, scale-up, stem cellphone assays, third-dimensional tradition, tissue engineering and cytotoxicity assays. special protocols for a wide selection of tools give you the middle of every bankruptcy, making new method simply available. everybody operating in organic and clinical learn, no matter if in academia or a advertisement association, working towards telephone tradition will profit tremendously from this e-book.
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Extra info for Animal Cell Culture: A Practical Approach 3rd Edition
USA, 53, 288. Barnes, D. and Sato, G. (1980). Anal. , 102, 255. Leibovitz, A. (1963). Am. J. , 78, 173. Peehl, D. M. and Ham, R. G. (1980). In Vitro, 16, 526. Stacey, G. , Masters, J. R. , Hay, R. , Drexler, H. , MacLeod, R. A. F. and Freshney, R. I. (2000). Cell contamination leads to inaccurate data: we must take action now. Nature, 356. K. 1 Introduction Small scale culture of cells in flasks of up to 1 litre volume (175 cm2 surface area) is the best means of establishing new cell lines in culture, for studying cell morphology and for comparing the effects of agents on growth and metabolism.
1 Cell viability The dye exclusion test is based on the concept that viable cells do not take up certain dyes, whereas dead cells are permeable to these dyes. 4%) is the most commonly used dye, but has the disadvantage of staining soluble protein. 4%) is often preferred. Cells are counted in the standard manner using a haemocytometer. Some caution should be used when interpreting results as the uptake of the dye is pHand concentration-dependent, and there are situations in which misleading results can be obtained.
G. glutamine), spatial restrictions, and mechanical/shear stress. As soon as one of these factors comes into effect, the culture is finished and the remaining resources of the system are wasted. The aim is, therefore, to delay the onset of any one factor until the accumulated effect causes cessation of growth, at which point the system has been maximally utilized. g. Hepes instead of bicarbonate), continuous gassing, generous headspace volume, enriched rather than basal media, with nutrient-sparing supplements such as non-essential amino acids or lactalbumin hydrolysate, perfusion loops through ultrafiltration membranes or dialysis tubing for detoxification (3) and oxygenation, and attention to culture and process design.