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The QIAGEN Guide to Animal Cell Culture

that pipetting does not create foam.

8. Proceed as required (e.g., with passaging, freezing, nucleic acid isolation, etc).

Table 1. Composition of solutions for animal cell culture protocols * Store 1x trypsin-EDTA solution at 20C. Small aliquots can be stored at 28C for 12 weeks. Work quickly when using trypsin during cell culture, since trypsin degrades and enzymatic activity declines at 37C.

Passaging cells

Many adherent cell cultures will cease proliferating once they become confluent (i.e., when they completely cover the surface of cell culture vessel), and some will die if they are left in a confluent state for too long. Adherent cell cultures therefore need to be routinely passaged, that is, once the cells are confluent, a fraction of the cells need to be transferred to a new cell culture vessel. Suspension cells will exhaust their culture medium very quickly once the cell density becomes too high, so these cultures similarly require regular passaging.

IMPORTANT: Although regular passaging is necessary to maintain animal cell cultures, the procedure is relatively stressful for adherent cells as they must be trypsinized. We do not recommend passaging adherent cell cultures more than once every 48 h.

1. Harvest the cells, either by trypsinization (adherent cell cultures) or by centrifugation at 200 x g for 5 min (suspension cell cultures). Resuspend the cells in an appropriate volume of prewarmed growth medium containing serum.

Tip- The volume of medium used to resuspend the cells depends on the spl
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