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

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. Here, we describe a three-dimensional (3D) approach to cell culture in vitro, utilizing a bioreactor system designed to control oxygenation of 3D cancer cell cultures, in order to better mimic tumor microenvironments observed in vivo. We found transcriptomic differences in breast and ovarian cancer cell cultures grown in traditional monolayer cultures as compared to cultures grown 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% O2 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. Overall design: Examination of the effects of oxygenation gradients on 3D cancer cell cultures grown in a bioreactor. 3D cell cultures grown in 3% oxygen, 21% oxygen, and gradient oxygen are examined and compared to cells grown in 2D 21% oxygen. MCF-7 and OVCAR-8 cell lines are investigated.

常用的单层细胞培养（monolayer cell cultures）体系，无法为细胞培养提供符合生理学相关特征的环境，具体体现在细胞-细胞结构、细胞外基质（extracellular matrix）组成，以及关键生长因子与氧等小分子的时空递送等层面。
本研究介绍一种体外（in vitro）三维（3D）细胞培养方法，通过专为调控3D癌细胞培养氧合状态设计的生物反应器（bioreactor）系统，以更精准地模拟体内（in vivo）真实肿瘤微环境。
本研究发现，相较于在基质胶（Matrigel）三维基质中培养的细胞，传统单层细胞培养的乳腺癌与卵巢癌细胞系存在转录组（transcriptome）层面的差异。
此外，本研究借助该生物反应器系统，分析了分别在21% O₂、3% O₂以及3%至0% O₂氧气梯度环境下培养的3D细胞的转录组。
通过精准调控氧气递送，本研究观察到三种氧环境下细胞生长形态与转录组调控模式均存在显著差异。
实验设计概述：探究生物反应器中培养的3D癌细胞对氧合梯度的响应。
本研究对分别在3%氧气、21%氧气以及梯度氧气环境下培养的3D细胞进行检测，并与2D单层（21%氧气）培养的细胞进行对比。
所涉及的细胞系包括MCF-7与OVCAR-8。