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Insight into oxygenation levels within 3D cell models and its impact on cell metabolism
Author(s) -
Papkovsky Dmitri B,
Carey Conn,
Morten Karl,
Dmitriev Ruslan I,
Hynes James
Publication year - 2013
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.27.1_supplement.lb799
Subject(s) - intracellular , glycolysis , oxygenation , biophysics , cellular metabolism , oxidative phosphorylation , oxygen , oxygen tension , cell metabolism , chemistry , metabolism , cell , microbiology and biotechnology , oxidative metabolism , biochemistry , biology , ecology , organic chemistry
Appreciation that typical culture conditions reflect nonphysiological, hyperoxic states for most cell types, has spurred a trend to the use of 3D models and controlled O2 conc. to better reflect the in vivo condition. Importance of cellular O2 conc. is underscored by its impact on processes as diverse as stem cell differentiation or tumour drug resistance development. While control of ambient O2 is easily achievable, measurement of oxygenation at the monolayer and analysis of the critical O2‐ sensitive processes has presented significant challenge through lack of robust quantitative tools. We present a novel multiplexed fluorescence method using dedicated O2‐sensitive probes, allowing robust quantitative measurement of O2 conc. in biological specimens. Both intracellular oxygen and glycolytic flux are measured in parallel across range of 3D models and the effect of applied oxygen tension on tissue oxygenation and cell metabolism examined. Data presented illustrates that O2 conc. within 3D structures can deviate significantly from the environmental oxygen conc., and this can impact significantly on the balance between oxidative and glycolytic ATP generating pathways. These new high throughput (plate reader) and high content (imaging) offer the potential for new and detailed insight into the impact of physiological oxygen and its perturbation on metabolism and intracellular signalling.