Isolation and culturing of primary human colonocytes

In our laboratory, human cells and tissues are used as experimental models to understand the pathogenesis of human diseases. A reproducible method for the isolation and culturing of colonocytes from human surgical specimens has been developed. This method involves the dissection of the colon specimen to expose the mucosa, cleaning of the mucosa surface, and digestion of the mucosa with a mixture of pronase and collagenase. The dissociated colonocytes are then quantified and cultured on collagen-coated plastic. Our procedure yields approx. 30–40 million viable colonocytes per gram of tissue, with a viability of approx. 90%. We believe that the primary colonocyte cultures can be used to study important human diseases such as Crohn's disease and ulcerative colitis.