Isolation of proliferative epithelial cells from the rat cecum and proximal colon

Abstract

A two-step technique, resulting in an enriched subpopulation of viable proliferative epithelial cells from the rat cecum and proximal colon is reported. Chelation of divalent cations combined with mild mechanical dissociation was initially used to obtain a single-cell suspension of heterogeneous cell types. Subsequent velocity sedimentation, using a linear isokinetic gradient, yielded a subpopulation of homogeneous viable epithelial cells with only a small number of goblet cells. These cells were mitotically active, as assessed by thymidine kinase and [ 3H]thymidine incorporation, and appeared to originate from the proliferative region of rat colonic crypts.