By Maureen A. Harrison
Mobile tradition ideas are precious to the fashionable researcher yet tough to hold out effectively. As a part of the sequence of Handbooks in functional Animal phone Biology, this quantity bargains a concise functional advisor to the fundamental necessities of the approach. Researchers new to mobile tradition will discover a transparent rationalization of the fundamental gear of a tissue tradition facility, together with tissue tradition media and sera. It describes equipment for turning out to be suspension and adhesion cultures, together with tips on how to shop cells and get ready basic cultures from cells. For these already culturing cells, the guide will act as a convenient connection with the elemental options. The essence of the ebook is to accommodate the generalities of mobilephone tradition and to offer a grab of the elemental strategies ahead of involvement in additional really good paintings within the box. excellent for an individual entering into tissue phone tradition suggestions or searching for a concise reference booklet.
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Extra resources for General Techniques of Cell Culture (Handbooks in Practical Animal Cell Biology)
0 |JiS/cm conductivity) (equivalent to double distilled water) or greater has in our experience proved satisfactory. Modern deionizers regularly produce water up to 18 megohm/cm quality, although there are some who consider that it is not necessarily an advantage to have such pure water for the preparation of cell culture media. Filtration equipment When filters are steam sterilized in their housings, there is always the possibility damage may occur to the membrane that might not be detectable to the eye.
If medium is prepared on a fairly regular basis, it is worth preparing concentrated stock solutions of the amino acids, vitamins, glutamine, Phenol Red and certain of the salts used in the BSS. The following is a protocol for the small-scale in-house production of the medium and reagents required for basic cell culture. Equipment The exact type and size of individual pieces of equipment used will depend on the size of the batch of medium being prepared. The following are the general types of equipment needed: Preparation vessel in which to mix the ingredients (usually beaker, flask or tank1) Stirrer (magnetic for smaller batches, plastic rod or overhead for larger batches) Source of purified water (see further under Quality Control) 38 • General techniques of cell culture Sterile storage bottles in which to dispense the completed medium Sterile filter housing/cartridge(s) with ancillary lines and clamps Sterile dispensing bell2 (for larger batches).
This method has been used in our laboratory for dispensing hundreds of litres of medium each week for several decades without any contamination problems. Other solutions Growing and subculturing mammalian cells will also require a selection of other solutions. 014/°C. 5 at room temperature (23 °C). Sterilize by filtration. Use at a level of 10-25 mM (10-25 ml/litre) (Cytotoxic concentration = 100 mM). Hanks's BSS. requires 10 mM, Hams El2 20 mM and Earles BSS 40 mM. A general satisfactory compromise is to use 20 mM in all media.