Characterization of conditions for the primary culture of human small intestinal epithelial cells

Intestinal epithelial cells (IECs) are important for many aspects of gut ph ysiology and pathology. Different approaches have been tried for the primar y culture of human IECs, with varying degrees of success, as apoptosis easi ly occurs. Our aim was to develop a method for primary culture of human IEC s from biopsy material. IECs and Lamina propria (LP) cells were liberated f rom duodenal biopsies obtained from subjects undergoing routine endoscopy f or clinical investigations, whose small bowel was macroscopically normal. I ECs were cultured on collagen membranes in a 12-well tissue culture cluster , with LP cells and allogeneic Epstein-Barr Virus (EBV)-transformed B lymph ocytes (allo-B cells) underneath, in the well. Cultured IECs were character ized by light and confocal microscopy. Cytokine levels in culture supernata nts were measured by ELISA. Cells showed the columnar morphology of IECs, e ven after several days in culture. Best results were obtained from IECs cul tured above both LP and allo-B cells. IECs did not form monolayers as do tr ansformed epithelial cell lines, but they did preserve their original cell- cell contacts. Analysis of culture supernatants showed that IL-10 was produ ced by IECs initially, but IL-1ra was produced by LP cells in the underlyin g wells with increasing time in culture. Very little IL-1 beta was produced from any cultures. These results show that IECs can be isolated and mainta ined in primary culture for a short while, which could open new possibiliti es for research using patient material instead of cell lines.