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Reporter cell activity within hydrogel constructs quantified from oxygen-independent bioluminescence

Paper ID Volume ID Publish Year Pages File Format Full-Text
5890 445 2014 13 PDF Available
Title
Reporter cell activity within hydrogel constructs quantified from oxygen-independent bioluminescence
Abstract

By providing a three-dimensional (3D) support to cells, hydrogels offer a more relevant in vivo tissue-like environment as compared to two-dimensional cell cultures. Hydrogels can be applied as screening platforms to investigate in 3D the role of biochemical and biophysical cues on cell behaviour using bioluminescent reporter cells. Gradients in oxygen concentration that result from the interplay between molecular transport and cell metabolism can however cause substantial variability in the observed bioluminescent reporter cell activity. To assess the influence of these oxygen gradients on the emitted bioluminescence for various hydrogel geometries, a combined experimental and modelling approach was implemented. We show that the applied model is able to predict oxygen gradient independent bioluminescent intensities which correlate better to the experimentally determined viable cell numbers, as compared to the experimentally measured bioluminescent intensities. By analysis of the bioluminescence reaction dynamics we obtained a quantitative description of cellular oxygen metabolism within the hydrogel, which was validated by direct measurements of oxygen concentration within the hydrogel. Bioluminescence peak intensities can therefore be used as a quantitative measurement of reporter cell activity within a hydrogel, but an unambiguous interpretation of these intensities requires a compensation for the influence of cell-induced oxygen gradients on the luciferase activity.

Keywords
Hydrogel; Bioluminescence; Spatial distribution; Model; Luciferase; Oxygen
First Page Preview
Reporter cell activity within hydrogel constructs quantified from oxygen-independent bioluminescence
Publisher
Database: Elsevier - ScienceDirect
Journal: Biomaterials - Volume 35, Issue 28, September 2014, Pages 8065–8077
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering