1. A system for exposing molecules and cells to biologically relevant and accurately controlled steady-state concentrations of nitric oxide and oxygen.
- Author
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Dendroulakis V, Russell BS, Elmquist CE, Trudel LJ, Wogan GN, Deen WM, and Dedon PC
- Subjects
- Cell Line, Tumor, Cell Survival physiology, Culture Media chemistry, Culture Media metabolism, Humans, Models, Biological, Nitric Oxide chemistry, Nitric Oxide metabolism, Oxygen chemistry, Oxygen metabolism, Cell Culture Techniques methods, Nitric Oxide administration & dosage, Oxygen administration & dosage
- Abstract
Nitric oxide (NO) plays key roles in cell signaling and physiology, with diverse functions mediated by NO concentrations varying over three orders-of-magnitude. In spite of this critical concentration dependence, current approaches to NO delivery in vitro result in biologically irrelevant and poorly controlled levels, with hyperoxic conditions imposed by ambient air. To solve these problems, we developed a system for controlled delivery of NO and O(2) over large concentration ranges to mimic biological conditions. Here we describe the fabrication, operation and calibration of the delivery system. We then describe applications for delivery of NO and O(2) into cell culture media, with a comparison of experimental results and predictions from mass transfer models that predict the steady-state levels of various NO-derived reactive species. We also determined that components of culture media do not affect the steady-state levels of NO or O(2) in the device. This system provides critical control of NO delivery for in vitro models of NO biology and chemistry., (Copyright © 2012 Elsevier Inc. All rights reserved.)
- Published
- 2012
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