Method of preparing isolated colonic epithelial cells (colonocytes) for metabolic studies

Abstract

Suspensions of isolated colonic epithelial cells (colonocytes) have been obtained from rats by incubating everted lengths of colon with EDTA at 37°C in Krebs-Henseleit saline from which calcium was omitted and containing 0.25% (w/v) bovine serum albumin. Measurements of oxygen consumption and lactate production by cell suspensions indicate that they are metabolically active for at least one hour. The method has been modified for the preparation of isolated epithelial cells from the human colon by including an enzyme digestion step and by increasing the concentration of EDTA to 10 mM. Human colonocytes have been obtained either from normal mucosa (ascending and descending colon) or from the mucosa in ulcerative colitis (descending colon). Oxygen consumption of human cell suspensions is lower than in the rat but in colonocytes from both species the rate is increased by glucose and by n-butyrate, a normal constituent of the colonic lumen. The method yields metabolically active cell suspensions from diseased colonic mucosa and promises to be of value for biochemical studies of ulcerative colitis.