This report describes preliminary results of reconstitution of an in vivo-like multi-layered placental barrier without porous membrane support. We used thin collagen membrane (10 μm in thickness, commercially available) as permeable support of placental barrier structure and cultured BeWo trophoblastic cells and HUVECs on either side of the membrane. Co-cultured cells deposited abundant basement membrane matrices that support cellular function, whereas the deposition was hardly observed on polymer-based porous membrane conventionally used in organ-on-chip study. These results suggest that the placental barrier-like multi-layered construct integrated with microfluidic channels may serve as a platform for transplacental nutrients transport and pharmacokinetics studies.