Figure 1.

Sealed hypoxic cell culture plate. For hypoxic cell culture conditions, JAWS II cells are seeded under normoxic conditions in degassed, HEPES-buffered complete IMDM (selected wells are additionally supplemented with CoCl₂). Moreover, an oxygen absorber and an oxygen sensor (AgelessEye) are added to the 6-well plate to establish and monitor the low oxygen environment, respectively. Two sterile 0.75 mm plastic spacers placed on top of the 6-well plate allow gas exchange between the different wells of the cell culture plate. After the cell culture plate is closed with the lid, it is placed in a vacuum bag, evacuated from ambient air, and sealed using a vacuum sealing machine. Finally, the sealed hypoxic cell culture plate is then placed in a standard incubator at 37 °C. When exposed to low oxygen (≤0.5% O₂ concentration), the oxygen sensor AgelessEye turns from blue to pink. This color change can be controlled/monitored throughout the cultivation of the cells.