TAILIEUCHUNG - Aseptic Technique for Cell Culture

This unit describes some of the ways that a laboratory can deal with the constant threat of microbial contamination in cell cultures. Microorganisms are ubiquitous. Bacteria can be isolated from nearly any surface including inanimate objects and human skin. Fungal spores and bits of vegetative hyphae drift into a laboratory from air conditioning ducts and open doors. Mycoplasma infections most frequently originate from improperly sterilized media or serum. | Aseptic Technique for Cell Culture UNIT This unit describes some of the ways that a laboratory can deal with the constant threat of microbial contamination in cell cultures. Microorganisms are ubiquitous. Bacteria can be isolated from nearly any surface including inanimate objects and human skin. Fungal spores and bits of vegetative hyphae drift into a laboratory from air conditioning ducts and open doors. Mycoplasma infections most frequently originate from improperly sterilized media or serum. At the risk of eliciting paranoia in the novice cell culture user who has no training in microbiological techniques the possibility for microbial contamination exists everywhere. Inherent with successful manipulation of cell cultures is the basic understanding that everything that comes into contact with the cells must be sterile or noncontaminating. This includes media glassware and instruments as well as the environment to which the cultures are briefly exposed during transfer procedures. Because cleaning up a contaminated culture is too frequently a disheartening and unsuccessful experience the best strategy is to employ procedures to prevent microbial contamination from occurring in the first place. This unit begins with a protocol on aseptic technique see Basic Protocol 1 . This catch-all term universally appears in any set of instructions pertaining to procedures in which noncontaminating conditions must be maintained. In reality aseptic technique cannot be presented in one easily outlined protocol but rather encompasses all aspects of environmental control personal hygiene equipment and media sterilization and associated quality control procedures needed to ensure that a procedure is indeed performed with aseptic noncontaminating technique. Although cell culture can theoretically be carried out on an open bench in a low-traffic area most cell culture work is carried out using a horizontal laminar-flow clean bench see Basic Protocol 2 or a vertical laminar-flow .

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