Abstract: Objective: To establish in vitro blood-brain barrier model. Methods: Rat brain microvessel endothelial cells and astrocytes were cultured and immunohistochemical stainning were employed to identify the cells. The two kinds of cell were then seeded onto each surface of polyester-multipore-membrane. Growth condition of the two kinds of cell on the membrane was evaluated by phase contrast microscope and HE stainning. Results: The two kinds of cultured cellpresent typical shape of brain microvessel endothelial cell and astrocyte respectively.Immunohistochemical stainning proved that the purity of the two kinds of cell was above 90% and 95%. The two kinds of cell both formed continued monolayer on each surface of the polyester-multipore-membrane. Conclusions: Coculturingbrain microvessel endothelial cells and astrocytes on polyester-multipore-membrane resemle in vivo blood-brain barrier closely, which can be used as in vitro blood-brain barrier model.