Two different co-culture methods, on coverslips and in Ibidi culture-inserts, were established in several test series. The main advantage of the co-culture in Ibidi culture-inserts is the confrontation of the endometrial epithelial cell lines and the extravillous trophoblast in a defined cell-free gap. In contrast to the co-cultures without Ibidi inserts it is ensured that every cell line can grow separately at the beginning of the co-culture. After removing the culture-insert the invasion of the trophoblast cells can take place in the defined gap.With both co-culture methods the interaction between the extravillous trophoblast cells and the endometrial epithelial cells could be simulated and analyzed.Using three different receptive endometrial epithelial cell lines HEC-1-A, Ishikawa and RL95-2, it could be shown that the invasive behaviour of the trophoblast cell line AC-1M88 in confrontation with these cell lines is different. The invasive behaviour of the trophoblast cells in confrontation with the weakly polarized RL95-2 cell line was stronger in comparison with the moderately polarized Ishikawa cell line and the highly polarized HEC-1-A cell line. The formation of desmosomal and adherens junction proteins between endometrial epithelial cells and trophoblast cells was different as well. The strongest expression of adhesion proteins was asserted between trophoblast cells and RL95-2 cells. Under the culture conditions in Ibidi inserts no formation of desmosomes and adherens junctions could be observed between HEC-1-A and AC-1M88 cells.Individual AC-1M88 cells expressed a stronger HLA-G signal than cells in a monolayer. Trophoblast cells in a monolayer with direct contact to another cell line showed a stronger HLA-G signal than cells without direct contact. The morphology of trophoblast cells changed in dependence of the endometrial epithelial cell line used in co-culture. Trophoblast cells displayed more distinct cellular protrusions in confrontation with the weakly polarized RL95-2 cell line than with the moderately polarized Ishikawa cell line and the strongly polarized HEC-1-A cell line.It could be shown that the invasive behaviour and the morphology of trophoblast cells as well as the adhesion by desmosomes and adherens junctions between endometrial epithelial and trophoblast cells is changed in dependence of the polarity of the different endometrial epithelial cell lines.