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  • A causal relation between bioluminescence and oxygen to quantify the cell niche.

A causal relation between bioluminescence and oxygen to quantify the cell niche.

PloS one (2014-05-21)
Dennis Lambrechts, Maarten Roeffaers, Karel Goossens, Johan Hofkens, Greetje Vande Velde, Tom Van de Putte, Jan Schrooten, Hans Van Oosterwyck
ABSTRACT

Bioluminescence imaging assays have become a widely integrated technique to quantify effectiveness of cell-based therapies by monitoring fate and survival of transplanted cells. To date these assays are still largely qualitative and often erroneous due to the complexity and dynamics of local micro-environments (niches) in which the cells reside. Here, we report, using a combined experimental and computational approach, on oxygen that besides being a critical niche component responsible for cellular energy metabolism and cell-fate commitment, also serves a primary role in regulating bioluminescent light kinetics. We demonstrate the potential of an oxygen dependent Michaelis-Menten relation in quantifying intrinsic bioluminescence intensities by resolving cell-associated oxygen gradients from bioluminescent light that is emitted from three-dimensional (3D) cell-seeded hydrogels. Furthermore, the experimental and computational data indicate a strong causal relation of oxygen concentration with emitted bioluminescence intensities. Altogether our approach demonstrates the importance of oxygen to evolve towards quantitative bioluminescence and holds great potential for future microscale measurement of oxygen tension in an easily accessible manner.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Tetraethyl orthosilicate, 99.999% trace metals basis
Sigma-Aldrich
Tetraethyl orthosilicate, ≥99.0% (GC)
Sigma-Aldrich
Tetraethyl orthosilicate, reagent grade, 98%
Sigma-Aldrich
Tetraethyl orthosilicate, packaged for use in deposition systems
Supelco
Rhodamine 6G, analytical standard
Sigma-Aldrich
Fluorescein, for fluorescence, free acid
Amphotericin B, European Pharmacopoeia (EP) Reference Standard
Fluorescein, European Pharmacopoeia (EP) Reference Standard