An atraumatic method to establish human colon carcinoma in long-term culture

Techniques for creation of colon carcinoma epithelial cells lines in long-t erm culture have been available for years, but these techniques have involv ed mechanical or enzymatic methods to separate epithelial cells from surrou nding tissues. While this practice has been intermittently successful, the effect of these traumatic methods on long-term cellular behavior is unknown . Samples of colon carcinoma from patient volunteers were subjected to seri al nonenzymatic disruptions of carcinoma cells from surrounding fibrous tis sues. Cells were collected, allowed to proliferate, and then tested for the ir epithelial characteristics (mucin, vimentin, cytokeratin, colon-specific antigen, carcinoembryonic antigen) by immunohistochemistry and flow cytome try. Growth characteristics were determined by phase-contrast microscopy, m ultiple passage, and freeze/thaw effects. Tumorigenicity was proven in nude mice. Of 11 initial attempts, three resulted in stable long-term culture l ines of cells which are demonstrated to behave similarly to the original tu mors hom which they were derived. This technique adds another reliable in v itro tool for the study of colon carcinoma, (C) 1999 Academic Press.