Isolation and characterization of epithelial cell types from the normal rat colon

Abstract

Populations enriched in proliferative, mucous goblet, and absorptive cells were isolated by a method of repeated time dissociation of the colonic mucosa from the adult rat. Five sequential cell suspensions were obtained by rotating the everted colon on 0.2% trypsin solution in Eagle's minimum essential medium at 30 degrees for 20 min each time. Approximately 3.7 x 10(7) cells were obtained from each colon, and 87% of the cells were found viable. The results, based on the [3H]-thymidine-labeling index, [3H]thymidine incorporation, thymidine kinase activity, and histochemical and ultrastructural observations, indicate the Cell Suspension I, separated from the luminal surface, contains 82 +/- 9% (S.D.) absorptive cells; Suspension III, separated from the middle level of the crypts, contains 80 +/- 7% mucous cells. Suspension V, isolated from the crypt base, contains the majority of proliferative epithelial cells (85 +/- 10%). This method provides a suitable tool for a variety of studies in colon carcinogenesis.