The establishment of an in vitro blood-brain barrier

Objective To establish a convenient and reliable method for research of blood-brain barrier (BBB) in vitro. Methods After coculturing of ECV304 with astrocytes, transendothelial electrical resistance of monolayer of ECV304 was determined and the presence of tight junctions between endothe-lial cells was investigated under an electrical microscope. The activity of γ-glutamyl transpeptidase (γ-GT) and alkaline phosphatase (ALP) were measured. Results The BBB in vitro has a similar ultrastruc-ture with BBB such as tight junctions between endothelial cells, lots of mitochondria and little pinocytosis. The in vitro BBB had a high TER value of 321.3Ωcm2 after coculture for 10 days. The activity of γ- GT and ALP of ECV304 were increased and could reach (480. 24 ± 104. 17) U/mg protein, (589. 13 ± 54.35) U/mg protein respectively after coculture with astrocytes. Conclusion Some structural and functional features of BBB can be induced in the blood-brain model. The method of coculture of ECV304 and astrocytes can be a useful model for research of BBB in vitro.