In vitro cell culture pO2 is significantly different from incubator pO2

Continuous noninvasive monitoring of peri-cellular liquid phase pO2 in adherent cultures is described. For neurons and astrocytes, this approach demonstrates that there is a significant difference between predicted and observed liquid phase pO2. Particularly at low gas phase pO2s, cell metabolism shifts liquid phase pO2 significantly lower than would be predicted from the O2 gas/air equilibrium coefficient, indicating that the cellular oxygen uptake rate exceeds the oxygen diffusion rate. The results demonstrate the need for direct pO2 measurements at the peri-cellular level, and question the widely adopted current practice of relying on setting the incubator gas phase level as means of controlling pericellular oxygen tension, particularly in static culture systems that are oxygen mass transfer limited.