Spatial control of oxygen delivery to 3D cultures alters cancer cell growth and gene expression

Abstract

Commonly used monolayer cancer cell cultures fail to provide a physiologically relevant environment in terms of oxygen delivery. Here, we describe a three-dimensional bioreactor system where cancer cells are grown in Matrigel in modified 6-well plates. Oxygen is delivered to the cultures through a polydimethylsiloxane (PDMS) membrane at the bottom of the wells, with microfabricated PDMS pillars to control oxygen delivery. The plates receive 3% oxygen from below and 0% oxygen at the top surface of the media, providing a gradient of 3% to 0% oxygen. We compared growth and transcriptional profiles for cancer cells grown in Matrigel in the bioreactor, 3D cultures grown in 21% oxygen, and cells grown in a standard hypoxia chamber at 3% oxygen. Additionally, we compared gene expression of conventional 2D monolayer culture and 3D Matrigel culture in 21% oxygen. We conclude that controlled oxygen delivery may provide a more physiologically relevant 3D system.

Spatial control of oxygen delivery to three‐dimensional cultures alters cancer cell growth and gene expression

“…Gene Ontology (GO) analysis of significantly, differentially regulated genes was performed with the GOseq package and redundant GO Biological processes were removed using REVIGO with “Small (0.5)” similarity and default parameters (Supek, Bosnjak, Skunca, & Smuc, ; Young, Wakefield, Smyth, & Oshlack, ). The raw data that form the basis of this study have been deposited in NCBI's Gene Expression Omnibus (Wulftange, ) and are accessible through GEO Series accession number GSE129111 (https://www.ncbi.nlm.nih.gov/geo/query/acc.cgi?acc=GSE129111).…”

Photo-Uncaging of a Microtubule-Targeted Rigidin Analogue in Hypoxic Cancer Cells and in a Xenograft Mouse Model

“…An important cellular effect of hypoxia is as a modulator of cell proliferation. 41 For many cell types, including A549, hypoxia was reported to induce decreased cell proliferation, 42,43 since an increase in the number of cells would result in increased oxygen demand and accompanying hypoxic stress. 41 MRC-5 fibroblasts are mostly responsible for extracellular matrix production in lungs.…”

Harnessing preclinical models for the interrogation of ovarian cancer

“…The underlying possibilities for this phenomenon are not only manifold but involute. Lack of cell-cell and cell-matrix interaction can be partly blamed while the diffusion rate of nutrients and metabolic waste might differ greatly in cell aggregates as well [125]. The structural difference in the culturing system brings genomic unconformity and the consequences are far more than that.…”