Cryopreservation Manual NucleoLink App. In some cases it is necessary to harvest cells culture vessels without using protein digesting enzymes, proteases, such as trypsin, collagenase and pronase. These proteases may modify cellular proteins or cell membranes, thus affecting some immunological or physiological assays.
Cells dissociate from each other as well as from plastic and glass.
Different cell lines may require different concentrations of EDTA and various incubation times. The addition of a complete growth medium or buffer containing balanced calcium and magnesium reconstitutes cells for re-seeding or assay procedures. As viability is dependent on how cells are handled and on the laboratory technique employed, these solutions are good alternatives when enzymes are undesirable. Pre-warm all reagents to 37 C. 2. Remove medium from the culture vessel without drying the monolayer.