TIME-RELATED EFFECTS OF ENZYMATIC DISAGGREGATION ON MODEL HUMAN LUNG CARCINOMAS

Enzyme cocktails used to prepare tumor cell suspensions may influence yield, viability, and cytology, thus time-related cocktail effects on model human lung carcinomas were examined. A549, NCI-H125, and NCI-H46 0 carcinomas were completely disaggregated at 25 degrees C over 2 h wi th either (mg/ml) collagenase/DNAse (C/D, 1/0.1), collagenase/hyaluron idase/DNAse (C/H/D, 1/0.1/0.1), or polymyxa protease/DNAse (PP/D, 3/0. 1). Trypan blue viabilities, total yields, viable yields, and now cyto metric percent tumor cells (TC) were measured every 20-30 min (n = 4-7 per tumor type). The final percentages of TC, mononuclear cells (MN), polymorphonuclear cells (PMN), lymphocytes, and necrotic cells were d etermined by cytology (n = 4-5 per tumor type). The time-dependent mea surements showed that 1) disaggregation was progressive and complete w ith all cocktails; 2) viability was stable or increasing with all cock tails; 3) percent TC was stable for all cocktails, but lower for PP/D than C/D in final suspensions; and 4) PP/D gave lower final total yiel ds, higher final viabilities, but the same final viable yields as the C cocktails, suggesting selective elimination of dead cells by PP/D. F inal cytology measurements showed that PP/D gave a lower percent MN an d a higher percent PMN than C cocktails. Cocktail effects may importan tly influence cell suspension properties. (C) 1995 Wiley-Liss, Inc.