Generating linear oxygen gradients across 3D cell cultures with block-layered oxygen controlled chips (BLOCCs)
| Autor: | Rachael M. Kenney, Matthew R. Lockett, Matthew W. Boyce, William C. Simke |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Fabrication
Materials science General Chemical Engineering Cell Culture Techniques Silicones chemistry.chemical_element Silicone rubber 01 natural sciences Oxygen Article Analytical Chemistry 03 medical and health sciences chemistry.chemical_compound Silicone Humans Hypoxia Tissue homeostasis 030304 developmental biology 0303 health sciences 010401 analytical chemistry General Engineering Hypoxia (environmental) 0104 chemical sciences Oxygen tension chemistry Biophysics Liquid bubble |
| Zdroj: | Anal Methods |
| ISSN: | 1759-9679 1759-9660 |
| DOI: | 10.1039/c9ay01690b |
| Popis: | Oxygen is a transcriptional regulator responsible for tissue homeostasis and maintenance. Studies relating cellular phenotype with oxygen tension often use hypoxia chambers, which expose cells to a single, static oxygen tension. Despite their ease of use, these chambers are unable to replicate the oxygen gradients found in healthy and diseased tissues. Microfabricated devices capable of imposing an oxygen gradient across tissue-like structures are a promising tool for these studies, as they can provide a high density of information in a single experimental setup. We describe the fabrication and characterization of a modular device, which leverages the gas-permeability of silicone to impose gradients of oxygen across cell-containing regions, assembled by layering sheets of laser cut acrylic and silicone rubber. The silicone also acts as a barrier, separating the flowing gases from the cell culture medium, preventing evaporation or bubble formation in experiments that require prolonged periods of incubation. The acrylic components provide a rigid framework to provide a sterile culture environment. Using oxygen-sensing films, we show the device can support gradients of different ranges and steepness by simply changing the composition of the gases flowing through the silicone components of the BLOCC. Using a cell-based reporter assay, we demonstrate that cellular responses to hypoxia are proportional to oxygen tension. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|