A three-zone hypoxia chamber capable of regulating unique oxygen and carbon dioxide partial pressures simultaneously

Oxygen is a vital but often overlooked variable in tissue culture experiments. Physiologically relevant oxygen tensions range from partial pressures of 100 mmHg at the alveolar-capillary interface in the lung to less than 7.6 mmHg in the hypoxic regions of solid tumors. These values are markedly lower than the partial oxygen pressure of ambient air, which is standard experimental practice. Physiologically relevant culture environments are needed to better predict cellular and tissue-level responses to drugs or potential toxins. Three commonly used methods to regulate in vitro oxygen tension involve placing cells in 1) a hypoxia chamber, 2) setups that rely on mass transport-limited microenvironments, and 3) microfabricated devices. Hypoxia chambers have the lowest barrier to entry, as they do not require laboratories to change their tissue culture setups. Here, we present a gas-regulation system for a three-zone hypoxia chamber. Each zone can maintain independent environments, with partial pressure compositions of 1-21 % O₂ and 1-10 % CO₂. The design incorporates small-scale fabrication techniques (e.g., laser cutting and 3D printing) and off-the-shelf electronic components for simple assembly. The hypoxia chambers are significantly lower in cost than the commercial counterparts: $1,400 for the control system or $4,100 for a complete three-zone chamber system.