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

Southern Research Institute, Birmingham, Alabama 35255

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 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 observations, 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.

¹ Supported by Contract NO1-CP-75914 from the National Cancer Institute.

² To whom requests for reprints should be addressed.

Received 6/18/79. Accepted 10/ 3/80.