Isolation and Characterization of Epithelial Cell Types from the Normal Rat Colon1

Populations enriched in proliferative, mucous goblet, and absorptive cells were isolated by a method of repeated time dissociation of the colonie mucosa from the adult rat. Five sequential cell suspensions were obtained by rotating the everted colon in 0.2% trypsin solution in Eagle's minimum essential medium at 30°for 20 min each time. Approximately 3.7 x 107 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 ob servations, indicate that 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.