A co-culture model of the two most abundant intestinal epithelial cell types (absorptive and goblet cells) was developed on permeable supports using the human Caco-2 and HT29-H cell lines. Seeding conditions were selected to give a proportion of goblet cells comparable to the 25-55% found in human colon. The co-cultures comprised small clusters of HT29-H goblet cells embedded in Caco-2 absorptive cells. Electron microscopy showed that tight junctions formed between the two cell types in co-culture. Mono-cultures of HT29-H goblet cells were more permeable to ions and hydrophilic marker molecules and peptides and had a lower Isc than mono-cultures of Caco-2 absorptive cells. The permeability of the co-cultures was intermediate between those of the two mono-cultures. The co-cultures provide a model where absorptive and goblet cells can be studied simultaneously in studies on e.g. drug binding and transport, as well as in studies on absorption enhancement.