Mixing and delivery of multiple controlled oxygen environments to a single multiwell culture plate.

Abstract

Precise oxygen control is critical to evaluating cell growth, molecular content, and stress response in cultured cells. We have designed, fabricated, and characterized a 96-well plate-based device that is capable of delivering eight static or dynamically changing oxygen environments to different rows on a single plate. The device incorporates a gas-mixing tree that combines two input gases to generate the eight gas mixtures that supply each row of the plate with a different gas atmosphere via a removable manifold. Using air and nitrogen as feed gases, a single 96-well plate can culture cells in applied gas atmospheres with Po2 levels ranging from 1 to 135 mmHg. Human cancer cell lines MCF-7, PANC-1, and Caco-2 were grown on a single plate under this range of oxygen levels. Only cells grown in wells exposed to Po2 ≤37 mmHg express the endogenous hypoxia markers hypoxia-inducible factor-1α and carbonic anhydrase IX. This design is amenable to multiwell plate-based molecular assays or drug dose-response studies in static or cycling hypoxia conditions.

• Authors: Ming Yao, Tyler Sattler, Zahid N Rabbani, Thomas Pulliam, Glenn Walker, Michael P Gamcsik
• Journal: American journal of physiology. Cell physiology (Am J Physiol Cell Physiol) Vol. 315 Issue 5 Pg. C766-C775 (11 01 2018) ISSN: 1522-1563 [Electronic] United States
• PMID: 30183322 (Publication Type: Journal Article, Research Support, N.I.H., Extramural)

Chemical References

• Hypoxia-Inducible Factor 1, alpha Subunit
• Oxygen

Topics

• Caco-2 Cells
• Cell Culture Techniques (instrumentation)
• Cell Hypoxia (genetics)
• Cell Proliferation (genetics)
• Humans
• Hypoxia-Inducible Factor 1, alpha Subunit (chemistry, metabolism)
• MCF-7 Cells
• Oxygen (chemistry, metabolism)