A new test to measure homotypic aggregation of human tumour cells

A method is described for quantitative measurements of homotypic aggregation by sequential passaging of cells through several gauze nets with different mesh width. This method allows rapid and simple determination of the size distribution of the formed aggregates with little cost. Time course and the effects of divalent cations, sugars and of enzyme treatment on homotypic aggregation were examined in detail for the human colon carcinoma line HT29, but also aggregation of human neuroblastoma, leukemic promyelocytes HL60, and of murine lymphoma cells was studied. Crude membrane fractions prepared from several colon carcinoma cells and from dissociated human colon tumour tissue showed strong aggregation-promoting effects when incubated with HT29 cells. Determination of lectin-induced agglutination of HT29 cells by means of the proposed method demonstrated that HT29 carries high numbers of binding sites for Ricinus communis agglutinin, wheat germ agglutinin, Ulex europeus agglutinin and Griffonia simplicifolia I isolectin A4. These results were supported by direct microanalytical determination of membrane-bound sialic acid and total fucose.