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finding is in agreement with other studies that have reported in vivo O2 content of the microvasculature. bEnd.3 cells cultured under 5 kPa also exhibited lower intracellular glutathione content compared to cells cultured under 18 kPa O2 (20.6 vs 29.9 mmol/mg protein, n=4, p=0.003). Furthermore, bEnd.3 cells cultured under 5 kPa O2 exhibited greater sensitivity to OGD-induced cell death, with increased ratio of dead to live cells seen under 2 hour OGD (0.42 vs 0.05, n=3, p=0.0002). Conclusions The majority of cell culture studies have conducted experiments under hyperoxia. The present study highlights the importance of conducting cell culture studies under physiological O2 levels, as demonstrated by the different phenotype resulted from long-term culture under physiological O2 levels.