*2.1.6 Simulating ischemia and reperfusion using cultured cells*

Cultured brain capillary endothelial cells were exposed to conditions simulating ischemia and reperfusion by incubating first at 37°C in an ischemic medium (without glucose, pH 6.8) equilibrated with an atmosphere of 95% N2 and 5% CO2, followed by simulated reperfusion in a control medium (5.6 mM glucose, pH 7.4) equilibrated with room air and 5% CO2 [38, 39]. To provide a constant environment, the cells were maintained in sealed chambers (Billups-Rothenberg, CA) pre-equilibrated to the desired atmospheric conditions [39].
