Rat enterocytes were cultured on human amniotic membranes.Intestine of neonatal DA rats was digested using collagenase and dispase according to the technique developed by Evans. The harvested enterocytes were cultured on human amniotic membranes using standard cell culture techniques.After the second day of culture, some intestinal epithelial units started to gradually detach from the membrane, dispersing as single cells and disappearing within a few days. On the contrary, other units showed signs of cell proliferation. The cultured cells underwent morphologic changes, survived, and remained attached to the amniotic membrane for 3 weeks. Paraffin sections of the membrane showed cultured cytokeratin-positive cells attached to the membrane as a monolayer.Human amniotic cell membranes help to maintain rat enterocytes in culture for a long time period (3 weeks), possibly via secretion of trophic factors. This technique may provide a valuable tool to study the development and the properties of these epithelial cells in a culture environment.