Isolation and Culture of Colon Fibroblasts

Fibroblasts were isolated from primary tissues using previously described methodologies [3 (link)]. Briefly, after mincing, heated collagenase digestion on a shaking water bath was used and the filtrate was cultured on tissue culture plates in DMEM 10% FBS (Atlanta Biologicals) with 1% Pen/Strep (Gibco). Human colon fibroblasts cell lines CRL1541, CRL1459, and CRL7213 were obtained from the American Type Culture Collection (ATCC). Primary colon fibroblasts were isolated from human colon tissue was obtained with informed consent from patients undergoing surgery without preoperative treatment for sporadic colon cancer or colectomy for chronic ulcerative colitis. Primary fibroblasts were cultured from non-dysplastic sections of colon (normal), adenocarcinoma or colitic sections as previously described [3 (link)].