Background: Colorectal tumors arise from unregulated cell proliferation of the intestinal epithelium through a multistep process the first step usually being premalignant adenomas. Familial adenomatous polyposis patients carry a germ line mutation in the APC gene leading to the development of thousands of polyps, which, if left untreated, lead to cancer. The goal of this study was the establishment of conditions for the culture of epithelial cells composing an adenomatic structure.
Materials and methods: All colorectal specimens were obtained from FAP patients within 1-2 hours of surgery. Cells were cultured by standard procedures. Characterization was carried out by immunostaining with pancytokeratin, vimentin and desmoplakin antibodies.
Results: A culture protocol that gave rise to epithelial cell growth with high efficiency and efficacy was established. Successful subculturing of the cell sheets took place only when dispase prepared in Ca2+ and Mg2+ free medium, was used to digest polyp tissue taken from FAP patients. By using immunostaining these cells were characterized as epithelial.
Conclusion: The protocol we developed here provides a means of preparing cell cultures from human colorectal adenomas, which aid in the research of the transition from adenoma to carcinoma.