96-Well Oxygen Control Using a 3D-Printed Device
| Autor: | Adam Szmelter, David T. Eddington, Jason Jacob |
|---|---|
| Rok vydání: | 2021 |
| Předmět: |
Gradient generator
3d printed Chemistry 010401 analytical chemistry Microfluidics Cell Culture Techniques Hypoxia (environmental) chemistry.chemical_element Equipment Design Microfluidic Analytical Techniques 010402 general chemistry 01 natural sciences Anticancer drug Oxygen Article 0104 chemical sciences Analytical Chemistry Volumetric flow rate A549 Cells Printing Three-Dimensional Humans Limiting oxygen concentration Single plate Biomedical engineering |
| Zdroj: | Anal Chem |
| ISSN: | 1520-6882 |
| Popis: | Oxygen concentration varies tremendously within the body and has proven to be a critical variable in cell differentiation, proliferation, and drug metabolism among many other physiological processes. Currently, researchers study the gas’s role in biology using low-throughput gas-control incubators or hypoxia chambers in which all cells in a vessel are exposed to a single oxygen concentration. Here, we introduce a device which can simultaneously deliver 12 unique oxygen concentrations to cells in a 96-well plate and seamlessly integrate into biomedical research workflows. The device inserts into 96-well plates and delivers gas to the headspace thus avoiding undesirable contact with media. This simple approach isolates each well using gas-tight pressure resistant gaskets effectively creating 96 “mini-incubators”. Each of the twelve columns of the plate is supplied by a distinct oxygen concentration from a gas-mixing gradient generator supplied by two feed gases. The wells within each column are then supplied by an equal flow-splitting distribution network. Using equal feed flow rates, concentrations ranging from 0.6% to 20.5% were generated within a single plate. A549 lung carcinoma cells were then used to show that O2 levels below 9% caused a stepwise increase in cell death for cells treated with the hypoxia-activated anti-cancer drug Tirapirizamine (TPZ). Additionally, the 96-well plate was further leveraged to simultaneously test multiple TPZ concentrations over an oxygen gradient and generate a 3D dose response landscape. The results presented here show how microfluidic technologies can be integrated into, rather than replace, ubiquitous biomedical labware allowing for increased throughput oxygen studies. |
| Databáze: | OpenAIRE |
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