Tissue disaggregation of @entity1 XXXX with further isopyknic and isokinetic gradient purification.

@entity1 @entity1358 @entity5
Five different @entity1 @entity1358 were disaggregated with three combinations of enzymes. Significant @entity5 heterogeneity in response to the enzyme disaggregation was observed. A combination of collagenase (0.5 mg/ml) and trypsin (0.25%) was then used for routine disaggregation of 11 additional @entity5 . The viability of cells in suspension ranged between 63 and 98% with a mean viability of 83.2 +/- 10.7% (S.D.). The mean yield of total viable cells per g of tissue was 17.4 +/- 14.2 x 10(6). @entity5 cells were further fractionated in isopyknic and isokinetic gradients. After isokinetic sedimentation, significant heterogeneity among @entity5 was seen, but lymphocytes were consistently located in Fraction 7 +/- 2, whereas @entity5 cells were predominantly in Fraction 22 +/- 1. Malignant epithelial cells were enriched to a 85.8 +/- 9.4% (range, 69.5 to 92.5%) purity by isokinetic gradient centrifugation. Lymphocytes could be successfully separated from @entity5 cells using an isopyknic gradient. Controlled rate freezing of cells provided material for repeated experiments while short-term tissue culture prior to cell separation increased the proportion of viable cells in the suspension. Disaggregation of @entity1 @entity1358 and separation of malignant cells from @entity5 lymphocytes provides the foundation for characterizing these @entity5 biochemically and for analyzing hormonal responsiveness and the immunological characteristics of these @entity5 in vitro.
