Modulating Temporal and Spatial Oxygenation over Adherent Cellular Cultures

Abstract

Oxygen is a key modulator of many cellular pathways, but current devices permitting in vitro oxygen modulation fail to meet the needs of biomedical research. A microfabricated insert for multiwell plates has been developed to more effectively control the temporal and spatial oxygen concentration to better model physiological phenomena found in vivo. The platform consists of a polydimethylsiloxane insert that nests into a standard multiwell plate and serves as a passive microfluidic gas network with a gas-permeable membrane aimed to modulate oxygen delivery to adherent cells. Equilibration time is on the order of minutes and a wide variety of oxygen profiles can be attained based on the device design, such as the cyclic profile achieved in this study, and even oxygen gradients to mimic those found in vivo. The proper biological consequences of the device's oxygen delivery were confirmed in cellular models via a proliferation assay and western analysis of the upregulation of hypoxia inducible transcription factor-1α. These experiments serve as a demonstration for the platform as a viable tool to increase experimental throughput and permit novel experimental possibilities in any biomedical research lab.

Three‐Dimensional Models of the Human Brain Development and Diseases

“…Several microfluidic devices have been developed over the last decade to generate oxygen gradients over cellular cultures by relying on the oxygen permeability of polydimethylsiloxane (PDMS). [71–76] Overall, researchers have investigated multiple approaches to model the ischemia stroke, however current microfluidic-based in vitro stroke models are still very challenging and further improvements are needed to faithfully recapitulate the disease pathology and understand the responses of brain cells following oxygen deprivation in stroke.…”

Microfluidics in Malignant Glioma Research and Precision Medicine

“…[160] One improvement over these cell culture platforms is the implementation of gas microchannels into the cell culture plates that could generate a hypoxic environment, but these devices were limited to use within 2D cell cultures. [161] In order to culture cells in a 3D microenvironment under hypoxic conditions, microfluidic platforms, containing gas channels positioned above and below the cell chamber, can be used to introduce the desired gas conditions adjacent to cell chambers to control oxygen concentration. [162] Hybrid PDMS and polycarbonate films have also been used in these devices, wherein the gas impermeable polycarbonate film is patterned above cell channels to reduce diffusion of oxygen from the atmosphere while PDMS enables oxygen diffusion from gas channels.…”

Methods to study the tumor microenvironment under controlled oxygen conditions

“…Modifications to standard laboratory tools such as the Boyden chamber [49] and well plates [50] have been developed to control oxygen tension. These structures, commonly composed of poly(dimethylsiloxane) (PDMS) and created via replica molding, provide gas channels above the cell culture chamber.…”