Advances in measuring cellular bioenergetics using extracellular flux

Ferrick, David A.; Neilson, Andy; Beeson, Craig C.

doi:10.1016/j.drudis.2007.12.008

Cited by 395 publications

(391 citation statements)

References 34 publications

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“…Basal OCR and ECAR were measured, as well as the changes in OCR caused by the addition of the metabolic inhibitors described above. Several parameters were deducted from the changes in OCR and mitochondrial reserve capacity was calculated by subtracting basal respiration from maximal respiratory capacity as described previously (14). A mirror plate was set up and treated identically in parallel with the assay plate.…”

Section: Extracellular Flux Assaymentioning

confidence: 99%

Sensitization of Glioblastoma Cells to Irradiation by Modulating the Glucose Metabolism

Shen

Hau

Joshi

et al. 2015

Molecular Cancer Therapeutics

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Because radiotherapy significantly increases median survival in patients with glioblastoma, the modulation of radiation resistance is of significant interest. High glycolytic states of tumor cells are known to correlate strongly with radioresistance; thus, the concept of metabolic targeting needs to be investigated in combination with radiotherapy. Metabolically, the elevated glycolysis in glioblastoma cells was observed postradiotherapy together with upregulated hypoxia-inducible factor (HIF)-1a and its target pyruvate dehydrogenase kinase 1 (PDK1). Dichloroacetate, a PDK inhibitor currently being used to treat lactic acidosis, can modify tumor metabolism by activating mitochondrial activity to force glycolytic tumor cells into oxidative phosphorylation. Dichloroacetate alone demonstrated modest antitumor effects in both in vitro and in vivo models of glioblastoma and has the ability to reverse the radiotherapy-induced glycolytic shift when given in combination. In vitro, an enhanced inhibition of clonogenicity of a panel of glioblastoma cells was observed when dichloroacetate was combined with radiotherapy. Further mechanistic investigation revealed that dichloroacetate sensitized glioblastoma cells to radiotherapy by inducing the cell-cycle arrest at the G 2 -M phase, reducing mitochondrial reserve capacity, and increasing the oxidative stress as well as DNA damage in glioblastoma cells together with radiotherapy. In vivo, the combinatorial treatment of dichloroacetate and radiotherapy improved the survival of orthotopic glioblastoma-bearing mice. In conclusion, this study provides the proof of concept that dichloroacetate can effectively sensitize glioblastoma cells to radiotherapy by modulating the metabolic state of tumor cells. These findings warrant further evaluation of the combination of dichloroacetate and radiotherapy in clinical trials.

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Section: Extracellular Flux Assaymentioning

confidence: 99%

Sensitization of Glioblastoma Cells to Irradiation by Modulating the Glucose Metabolism

Shen

Hau

Joshi

et al. 2015

Molecular Cancer Therapeutics

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“…6 Traditionally, this method was conducted using a Clark electrode, 7 which lacks the throughput required for screening a large number of compounds on a daily basis. The recently developed multiwell, plate-based Seahorse XF96 Extracellular Flux Analyzer 8 and dual-parameter beadbased MitoXpress 9 assay address the need for more flexible and higher-throughput detection technologies in adherent cells in culture. However, the MitoXpress bead-based assay system has limitations of collecting data at 30 °C, a nonphysiological temperature; data analysis in kinetic mode was not suitable for screening applications; and there was the potential to miss uncouplers in end-point mode.…”

Section: Introductionmentioning

confidence: 99%

The Acute Extracellular Flux (XF) Assay to Assess Compound Effects on Mitochondrial Function

et al. 2015

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Numerous investigations have linked mitochondrial dysfunction to adverse health outcomes and drug-induced toxicity. The pharmaceutical industry is challenged with identifying mitochondrial liabilities earlier in drug development and thereby reducing late-stage attrition. Consequently, there is a demand for reliable, higher-throughput screening methods for assessing the impact of drug candidates on mitochondrial function. The extracellular flux (XF) assay described here is a plate-based method in which galactose-conditioned HepG2 cells were acutely exposed to test compounds, then realtime changes in the oxygen consumption rate and extracellular acidification rate were simultaneously measured using a Seahorse Bioscience XF-96 analyzer. The acute XF assay was validated using marketed drugs known to modulate mitochondrial function, and data analysis was automated using a spline curve fitting model developed at GlaxoSmithKline. We demonstrate that the acute XF assay is a robust, sensitive screening platform for evaluating drug-induced effects on mitochondrial activity in whole cells.

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“…[5][6][7] More recently, the fluorescence and phosphorescence O 2 sensors have permitted relatively simple and accurate monitoring of respiration in clinical samples of small quantities. [8][9][10][11] Measurements of mitochondrial respiration in digitonin-permeabilised fibroblasts and in isolated mitochondria from muscle specimens using phosphorescent microplates have been reported. 8 Other analytic instruments for assessing cellular bioenergetics are also described.…”

Section: Discussionmentioning

confidence: 99%

“…[4][5][6][7] More novel approaches for monitoring cellular bioenergetics have been reported recently. [8][9][10] Al-Jasmi et al described the use of a phosphorescence oxygen analyser to measure lymphocyte respiration in patients. 10 Their method was based on previously published principles.…”

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confidence: 99%

Mitochondrial Oxygen Consumption by the Foreskin and its Fibroblast Rich Culture = إستهلاك الأوكسجين في المايتوكوندريا من قبل القلفة و الخلايا الليفية

Al-Jasmi¹,

Pramathan²,

Swid³

et al. 2013

SQUMJ

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abstract:Objectives: This study investigated the feasibility of using a phosphorescence oxygen analyser to measure cellular respiration (mitochondrial O 2 consumption) in foreskin samples and their fibroblast-rich cultures. Methods: Foreskin specimens from normal infants were collected immediately after circumcision and processed for measuring cellular respiration and for culture. Cellular mitochondrial O 2 consumption was determined as a function of time from the phosphorescence decay of the Pd (II) meso-tetra-(4-sulfonatophenyl)-tetrabenzoporphyrin. Results: In sealed vials containing a foreskin specimen and glucose, O 2 concentration decreased linearly with time, confirming the zero-order kinetics of O 2 consumption by cytochrome oxidase. Cyanide inhibited O 2 consumption, confirming that the oxidation occurred mainly in the mitochondrial respiratory chain. The rate of foreskin respiration (mean ± SD) was 0.074 ± 0.02 μM O 2 min -1 mg -1 (n = 23). The corresponding rate for fibroblast-rich cultures was 9.84 ± 2.43 μM O 2 min -1 per 10 7 cells (n = 15). Fibroblast respiration was significantly lower in a male infant with dihydrolipoamide dehydrogenase gene mutations, but normalised with the addition of thiamine or carnitine. Conclusion: The foreskin and its fibroblast-rich culture are suitable for assessment of cellular respiration. However, the clinical utility of foreskin specimens to detect disorders of impaired cellular bioenergetics requires further investigation. Keywords

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Advances in measuring cellular bioenergetics using extracellular flux

Cited by 395 publications

References 34 publications

Sensitization of Glioblastoma Cells to Irradiation by Modulating the Glucose Metabolism

Sensitization of Glioblastoma Cells to Irradiation by Modulating the Glucose Metabolism

The Acute Extracellular Flux (XF) Assay to Assess Compound Effects on Mitochondrial Function

Mitochondrial Oxygen Consumption by the Foreskin and its Fibroblast Rich Culture = إستهلاك الأوكسجين في المايتوكوندريا من قبل القلفة و الخلايا الليفية

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