Cell biology international reports. vol:5 issue:2 pages:125-32
The dissociation of human fibroblast cultures with the bacterial neutral protease (Dispase II) was monitored by viability and growth measurement and was compared to the effect of trypsin and EDTA. Cell suspensions with high viability and excellent growth were obtained after 10 min incubation at 37 degrees C in 4 U/ml dispase in 0.02% EDTA. A two to three-fold increase in mitotic index occurred in the cultures within 48 h after dispase dissociation. The initial rate of aggregation was comparable to that of trypsin or EDTA dissociated cells, but attachment to a substratum and agglutination by Wheat Germ Agglutinin were markedly enhanced. The results indicate that dispase-EDTA provides a valuable alternative to the enzymatic dissociation with trypsin. Moreover, it is an additional tool for the dissociation of cultured cells and for the study of the surface properties of single cells.