resolution respirometer chamber  (Oroboros Instruments)

Journal: Communications Biology

Article Title: Myocardial disarray drives metabolic inefficiency in human cardiomyocytes

doi: 10.1038/s42003-025-08842-x

Figure Lengend Snippet: A Real-time oxygen consumption rate (OCR) measurements of micropatterned and non-patterned hiPSC-CMs by Seahorse extracellular flux analyzer. Cells were treated with the ATP synthase inhibitor oligomycin, the respiratory uncoupler FCCP, and the respiratory chain blockers rotenone and antimycin A. B Scatter plots show basal respiration, production of ATP, and maximal respiration. N = 8–11 differentiations. C Real-time extracellular acidification rate (ECAR) measurements of hiPSC-CMs non-patterned and micropatterned by Seahorse extracellular flux analyzer. Cells were treated with glucose, ATP synthase inhibitor oligomycin, and glucose analog 2-DG. D Scatter plots show glycolysis, non-glycolytic acidification, and glycolytic capacity. N = 4–12 differentiations. E Cell energy diagram showing normalized oxygen consumption rate (OCR) and normalized extracellular acidification rate (ECAR) of micropatterned and non-patterned hiPSC-CMS at the basal (circle points) and stressed (FCCP exposed, square points) conditions. F Representative profiles of oxygen consumption of non-patterned (top) and micropatterned (bottom) hiPSC-CMs measured in permeabilized cells with complex protocol using Oxygraph-2k high resolution respirometry (generated by DatLab). Digitonin (Dig, 10 µg/ml), pyruvate (P, 1 mM) and malate (M, 4 mM), ADP (2 mM), succinate (10 mM), amytal (2 mM), oligomycin (10 µM), and TMPD (0.5 mM) combined with ascorbate (2 mM) were added sequentially. Blue line stands for decreasing oxygen concentration in the oxygraphy chamber containing permeabilized cells. Red line stands for the changes of oxygen flux per volume. G Oxygen flux (attomol/sec/cell) quantified in non-patterned and micropatterned hiPSC-CMs. n = 7 in each group from 3 independent differentiation. Results, mean ± standard deviation. Statistical analyses: Mann–Whitney was used for B and D , two-way ANOVA was used for G. * p < 0.05, ** p < 0.01, ns: not significant for the indicated comparisons.

Article Snippet: Mitochondrial respiration was measured in digitonin-permeabilized hiPSC-CMs from micropatterned or non-patterned conditions (day 42) in an Oxygraph high-resolution respirometer chamber (Oroboros Oxygraph-2k, Innsbruck, Austria).

Techniques: Generated, Concentration Assay, Standard Deviation, MANN-WHITNEY

Journal: bioRxiv

Article Title: Myocardial disarray drives metabolic inefficiency in human cardiomyocytes

doi: 10.1101/2024.09.26.615170

Figure Lengend Snippet: A . Real-time oxygen consumption rate (OCR) measurements of micropatterned and non-patterned hiPSC-CMs by Seahorse extracellular flux analyzer. Cells were treated with the ATP synthase inhibitor oligomycin, the respiratory uncoupler FCCP, and the respiratory chain blockers rotenone and antimycin A. B . Scatter plots show basal respiration, production of ATP, and maximal respiration. N = 8-11 differentiations. C . Real-time extracellular acidification rate (ECAR) measurements of hiPSC-CMs non-patterned and micropatterned by Seahorse extracellular flux analyzer. Cells were treated with glucose, ATP synthase inhibitor oligomycin, and glucose analog 2-DG. D . Scatter plots show glycolysis, non-glycolytic acidification, and glycolytic capacity. N = 4-12 differentiations. E . Cell energy diagram showing normalized oxygen consumption rate (OCR) and normalized extracellular acidification rate (ECAR) of micropatterned and non-patterned hiPSC-CMS at the basal (circle points) and stressed (square points) conditions. F . Representative profiles of oxygen consumption of non-patterned (top) and micropatterned (bottom) hiPSC-CMs measured in permeabilized cells with complex protocol using Oxygraph-2k high resolution respirometry. Digitonin (Dig, 10 µg/ml), pyruvate (P, 1 mM) and malate (M, 4 mM), ADP (2 mM), succinate (10 mM), amytal (2 mM), oligomycin (10 µM), and TMPD (0.5 mM) combined with ascorbate (2 mM) were added sequentially. Blue line stands for decreasing oxygen concentration in the oxygraphy chamber containing permeabilized cells. Red line stands for the changes of oxygen flux per volume. G . Oxygen flux (attomol/sec/cell) quantified in non-patterned and micropatterned hiPSC-CMs. n = 7 in each group from 3 independent differentiation. Results, mean ± standard deviation. Statistical analyses: Mann-Whitney was used for B, and D, two-way ANOVA was used for G. * p<0.05, ** p<0.01, ns: not significant for the indicated comparisons.

Article Snippet: Mitochondrial respiration was measured in digitonin-permeabilized hiPSC-CMs from micropatterned or non-patterned conditions (day 42) in an Oxygraph high-resolution respirometer chamber (Oroboros Oxygraph-2k, Innsbruck, Austria).

Techniques: Concentration Assay, Standard Deviation, MANN-WHITNEY