DUCT EPITHELIAL-CELLS CULTURED FROM HUMAN PANCREAS PROCESSED FOR TRANSPLANTATION RETAIN DIFFERENTIATED DUCTAL CHARACTERISTICS

A procedure is described for the isolation and growth in vitro of epit helial cells from the duct network of human pancreas, referred to as D EC. A significant advantage of our procedure over previously published procedures is that it enables the isolation of DEC from small pieces of pancreas tissue (<5 g) and, also, from the digest remaining after t he isolation of islet cells from human pancreas, material that would n ormally be discarded. These were the only reliable sources for pancrea s tissue available to us. This procedure shows that some of the techni ques that have been successfully used for the isolation of rodent DEC are also valuable in the isolation of human DEC. In particular, the us e of cholera toxin to prevent fibroblast growth and contamination obvi ates the need for the time-consuming procedure of physically removing fibroblasts or the use of expensive fibroblast-specific monoclonal ant ibodies. The use of sieving to separate the digest immediately achieve s a partial purification, which, coupled with that of allowing duct cy sts to form, adds to the purity of the final preparation. The ductal s ystem of the intact pancreas tissue and the DEC derived from it expres sed cytokeratins 7, 8/18, and 19 and markers for the presence of MUC1, CFTR, and carbonic anhydrase II, which are specific for ductal epithe lial cells or for pancreatic ductal functions. This study showed that it is possible to obtain selectively viable DEC from small ducts in ot herwise waste pieces of human pancreas. It showed that these cells ret ained all of the epithelial characteristics that were examined and, in combination with data from an earlier study, showed that the cultured DEC retain the metabolic functions of duct epithelial cells in vivo.