CULTURE OF HUMAN MAIN PANCREATIC DUCT EPITHELIAL-CELLS

Attempts to grow human pancreatic duct epithelial cells in long-term c ulture have proven difficult. me have developed a system of growing th ese cells for several passages by adapting methods used to culture dog pancreatic duct cells. Epithelial cells were enzymatically dissociate d from the main pancreatic duct and plated onto collagen-coated cultur e inserts suspended above a human fibroblast feeder layer. After prima ry culture, the cells were either passaged onto new inserts or plastic tissue culture plates in the absence of collagen. Cells grown on the latter plates were maintained in a serum-free medium. Primary pancreat ic duct epithelial cells grow steadily to confluence as a monolayer in the feeder layer system. After primary culture, cells passaged onto n ew inserts with fresh feeder layer or plastic plates and fed with seru m-free medium continued to develop into confluent monolayers for up to four passages. The cells were columnar with prominent apical microvil li, sub-apical secretory vesicles, and lateral intercellular junctions resembling the morphology of normal in vivo epithelial cells. These c ells were also positive for cytokeratin 19, 7, and 8 and carbonic anhy drase II, as measured by immunohistochemistry. Metabolically, these ce lls synthesized and secreted mucin, as measured by incorporation of tr itiated N-acetyl-D-glucosamine. In conclusion, we demonstrated that hu man pancreatic epithelial cells from the main duct can be successfully grown in culture and repeatedly passaged using a feeder layer system, with serum-free medium, and in organotypic cultures.