Development of In Vitro Blood-Brain Barrier Model with Bovine Brain Microvessel Endothelial Cells / 체질인류학회지

ABSTRACT

The in vitro blood-brain barrier (BBB) model was established with bovine brain microvessel endothelial cells (BBMECs). The characteristics of BBMECs were identified by morphological and functional studies. BBMECs began to grow as spindle shaped cells and distinctly formed the monolayer of whirling appearance by 6 to 7 days after plating. Transendothelial electrical resistance (TEER) of the monolayer increased through 11 days and then started to decrease. The gamma-GTP activity (1791.5+/-397.8 nmol/min/mg of protein) and alkaline phosphatase activity (15.0+/-7.8 micromol/min/mg of protein) were high. BBMECs in culture were characterized by the binding of anti-vWF, anti-ZO-1, anti-vimentin, and anti-fibronectin antibodies. They failed to react with anti-GFAP, anti-GalC, and antineurofilament 160/200 kD antibodies, markers for astrocyte, oligodendrocyte, and neuron, respectively. Decreasing order of the permeability through the BBMEC monolayers of paracellular transport model drugs was mannitol, sucrose, and PEG-4000. The permeability of transcellular transport model drugs, progesterone and propranolol was much higher than that of paracellular transport model drugs. The BBMECs cultured on porous membrane have been qualified as an in vitro BBB model and also can be used for the BBB transport study in the drug development and for the BBB transport mechanism of drugs.