Bioreactor mimicry of blood vessel oxygen delivery alters cancer cell growth and gene expression patterns

Authors

• MM Gottesman
• WJ Wulftange
• M Garmendia-Cedillos
• NY Morgan
• TJ Pohida
• RW Robey

Abstract

The Laboratory of Cell Biology has a longstanding interest in developing models to study drug resistance mechanisms, including intrinsic and acquired resistance to anti-cancer drugs. While two-dimensional (2D) cell culture has led to remarkable advances in the study of cancer, cancer cells grown in monolayer frequently do not reflect the intrinsic drug resistance observed in tumors in vivo. Additionally, commonly used monolayer cell cultures lack the capacity to provide a physiologically relevant environment for cell culture in terms of cell-cell architecture, extracellular matrix composition, and spatiotemporal delivery of key growth factors and small molecules, such as oxygen. We have developed a bioreactor allowing moderately high-throughput measurements on 3D cultures maintained in controllably hypoxic or normoxic conditions, with gradients of oxygen concentration similar to those found in tumors. To accomplish this, an oxygen-transmissive membrane is patterned with a micropillar array, with the pillar spacing similar to typical intercapillary distances. We found transcriptomic differences in breast and ovarian cancer cell cultures grown in traditional monolayer cultures as compared to cultures grown in the bioreactor in a Matrigel three-dimensional matrix. We also investigated the transcriptomes of 3D cultures grown in 21% O2, 3% O2, and a gradient of 3% to 0% O2 using our bioreactor system. By controlling oxygen delivery, we observed differences in cell growth morphology and transcriptome regulation under the three conditions. Our work suggests that not only is the atmospheric concentration of oxygen relevant in cell culture, but also how it is delivered.

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