Cell fractionation of large bowel cancer

The fractionation of cells from human colonic carcinoma, a transplantable colonic carcinoma of the rat, and normal rat colonic mumsa by centrifugation in linear gradients of Ficoll (polysucrose) in tissue culture medium is described. Epithelial cells from human colonic carcinoma showed strong histochemical N-acetyl-@-D-glucosaminidase activity relative to stromal cells. The histochemical demonstration of this enzyme was employed to monitor the separation of malignant cells from other types of cells in human colonic carcinomas. After centrifugation, the cells from the quarter of the density gradient containing the highest percentage of cells with histochemically demonstrable N-acetyl-@-D-glucosaminidase also contained the highest levels of CEA/cell. Cells from this zone of the gradient gave rise to colonies when cultured in soft agar. Suspensions of cells from human colonic carcinomas contained eosinophils, plasma cells, lymphocytes, red blood cells and several unidentified morphological variants, which we believe to be of an epithelial nature. After centrifugation these morphological variants were located in the zone of the gradient that gave rise to colonies when cultured in soft agar. Similar morphological varieties were observed in both normal rat colonic mucosa and the transplantable rat tumor. The advantages of separated cells for a) the preparation of colonic cells for tissue culture and b) biochemical characterizations of malignant cells are described. Canc~r 40~2479-2486, 1977.