The Effect of Hypoxic and Normoxic Culturing Conditions in Different Breast Cancer 3D Model Systems

Abstract

The field of 3D cell cultures is currently emerging, and material development is essential in striving toward mimicking the microenvironment of a native tissue. By using the response of reporter cells to a 3D environment, a comparison between materials can be assessed, allowing optimization of material composition and microenvironment. Of particular interest, the response can be different in a normoxic and hypoxic culturing conditions, which in turn may alter the conclusion regarding a successful recreation of the microenvironment. This study aimed at determining the role of such environments to the conclusion of a better resembling cell culture model to native tissue. Here, the breast cancer cell line MCF7 was cultured in normoxic and hypoxic conditions on patient-derived scaffolds and compared at mRNA and protein levels to cells cultured on 3D printed scaffolds, Matrigel, and conventional 2D plastics. Specifically, a wide range of mRNA targets (40), identified as being regulated upon hypoxia and traditional markers for cell traits (cancer stem cells, epithelial–mesenchymal transition, pluripotency, proliferation, and differentiation), were used together with a selection of corresponding protein targets. 3D cultured cells were vastly different to 2D cultured cells in gene expression and protein levels on the majority of the selected targets in both normoxic and hypoxic culturing conditions. By comparing Matrigel and 3DPS-cultured cells to cells cultured on patient-derived scffolds, differences were also noted along all categories of mRNA targets while specifically for the GLUT3 protein. Overall, cells cultured on patient-derived scaffolds closely resembled cells cultured on 3D printed scaffolds, contrasting 2D and Matrigel-cultured cells, regardless of a normoxic or hypoxic culturing condition. Thus, these data support the use of either a normoxic or hypoxic culturing condition in assays using native tissues as a blueprint to optimize material composition.

Polysaccharides and Structural Proteins as Components in Three-Dimensional Scaffolds for Breast Cancer Tissue Models: A Review

“…Three-dimensional printing and bioprinting are other strategies to produce 3D scaffolds as they have the advantage of tuning the shape and properties of the scaffold as well as incorporating cells within the structure when mixed with the ink before printing. Three-dimensional printed scaffolds can mimic human tumor tissue more closely compared with 2D cell cultures [ 24 , 25 , 26 ].…”

Fast cycling of intermittent hypoxia in a physiomimetic 3D environment: A novel tool for the study of the parenchymal effects of sleep apnea

“…In conclusion, we have characterized an experimental approach, based on existing and easily available tools, that allows for controlled and precise application of high-frequency intermittent hypoxia both to 3D cell culture scaffolds such as ECM hydrogels as well as to precision-cut tissue slices. This approach, which is an advance on previous models investigating continuous hypoxia (Svanström et al, 2021), opens the door for the study of the specific noxious effects of intermittent hypoxia in the 3D-residing cells of the organs and tissues of patients with OSA under physiomimetic conditions. experimental support.…”