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An innovative intermittent hypoxia model in cell cultures allowing fast PO2 oscillations with minimal gas consumption

Minoves M, Morand J, Perriot F, Chatard M, Gonthier B, Lemarié E, Menut JB, Polak J, Pépin JL, Godin-Ribuot D, and Briançon-Marjollet A. Am J Physiol Cell Physiol. 2017 Jul 26:ajpcell.00098.2017. doi: 10.1152/ajpcell.00098.2017

Performing hypoxia-reoxygenation cycles in cell culture with a cycle duration accurately reflecting what occurs in obstructive sleep apnea (OSA) patients is a difficult but crucial technical challenge. Our goal was to develop a novel device to expose multiple cell culture dishes to intermittent hypoxia (IH) cycles relevant to OSA with limited gas consumption.

With gas flows as low as 200 mL/min, our combination of plate holders with gas-permeable cultureware generates rapid normoxia-hypoxia cycles. Cycles alternating 1 minute at 20% O2 followed by 1 minute at 2% O2 result in pO2 values ranging from 124 to 44mmHg. Extending hypoxic and normoxic phases to 10 minutes allows pO2 variations from 120 to 25mmHg. The volume of culture medium or the presence of cells only modestly affected the pO2 variations. In contrast, the nadir of the hypoxia phase increased when measured at different heights above the membrane. We validated the physiological relevance of this model by showing that HIF-1? expression was significantly increased by IH exposure in human aortic endothelial cells (HAoEC), murine breast carcinoma (4T1) cells as well as in a blood-brain barrier model (2.5, 1.5 and 6-fold increases, respectively).

In conclusion, we have established a new device to perform rapid intermittent hypoxia cycles in cell cultures, with minimal gas consumption and the possibility to expose several culture dishes simultaneously. This device will allow functional studies of the consequences of IH and deciphering of the molecular biology of IH at the cellular level using oxygen cycles that are clinically relevant to OSA.

Published on August 23, 2018