PRIMARY CULTURE AND TRANSFECTION OF EPITHELIAL-CELLS OF HUMAN SMALL-INTESTINE

Background: So far, no techniques are available for primary culture an d efficient transfection of human small-intestinal enterocytes, which would provide a valuable tool to investigate intestinal function. Meth ods: Human small-intestinal biopsy specimens were treated with collage nase and dispase. Resulting crypt units were cultured for several days . Using the intestinal epithelial cell lines Caco-2 and HT-29, we esta blished optimal conditions for transfection of a control plasmid, whic h were then applied to primary cultured cells. Results: Cells growing out of crypt units formed monolayer-like sheets and proliferated for s everal days. Most of the cells could be stained with antibodies agains t epithelial markers. Among seven different transfection reagents test ed, Lipofectamine was the most potent, with transfection efficiencies up to 25% for primary enterocytes. Conclusions: An easy technique was developed providing viable small-intestinal enterocytes that can be ef ficiently transfected.