BIOENERGETIC CHARACTERIZATION OF H9C2 CELLS USING THE EXTRACELLULAR FLUX ANALYZER

Authors

  • Ileana SCURTU University of Medicine and Pharmacy “Victor Babeș” - Timișoara
  • A. STURZU University of Medicine and Pharmacy “Victor Babeș” - Timișoara
  • Ioana Zinuca PAVEL University of Medicine and Pharmacy “Victor Babeș” - Timișoara
  • Roxana POPESCU University of Medicine and Pharmacy “Victor Babeș” - Timișoara
  • Oana Maria DUICU University of Medicine and Pharmacy “Victor Babeș” - Timișoara
  • Danina Mirela MUNTEAN University of Medicine and Pharmacy “Victor Babeș” - Timișoara

Abstract

The aim of the present work was to standardize the working methodology for assessing the bioenergetic profile of H9c2 cardiomyoblasts cells, with reference to the optimization of cell seeding number and the establishment of favorable concentrations for the classic modulators of mitochondrial respiratory function, in particular the one of a classical uncoupler, FCCP. Material and methods: The extracellular flux analyzer (XF, Seahorse Bioscience) is a novel high-throughput instrument able to monitor the metabolism of living cells by simultaneously measuring mitochondrial respiration and glycolysis. The in vitro platform will be further used to better understand the pathophysiology and the unrecognized side effects of drugs currently used in the therapy of major cardiovascular diseases. Conclusions: In the long run, characterization of novel pharmacological agents’ effects on other cell lines, including tumoral ones, will be also considered.

Author Biographies

  • Ileana SCURTU, University of Medicine and Pharmacy “Victor Babeș” - Timișoara

    Faculty of Medicine
    Department of Functional Sciences - Pathophysiology

  • A. STURZU, University of Medicine and Pharmacy “Victor Babeș” - Timișoara

    Faculty of Medicine
    Department of Functional Sciences - Pathophysiology

  • Ioana Zinuca PAVEL, University of Medicine and Pharmacy “Victor Babeș” - Timișoara

    Faculty of Medicine
    Department of Functional Sciences - Pathophysiology
    Department of Pharmacognosy

  • Roxana POPESCU, University of Medicine and Pharmacy “Victor Babeș” - Timișoara

    Faculty of Medicine
    Department of Morphological Sciences - Cellular and Molecular Biology

  • Oana Maria DUICU, University of Medicine and Pharmacy “Victor Babeș” - Timișoara

    Faculty of Medicine
    Department of Functional Sciences - Pathophysiology

  • Danina Mirela MUNTEAN, University of Medicine and Pharmacy “Victor Babeș” - Timișoara

    Faculty of Medicine
    Department of Functional Sciences - Pathophysiology

References

1. Chacko BK et al. The Bioenergetic Health Index: a new concept in mitochondrial translational research. Clin Soc 2014; 127(6): 367-373.
2. Blake R and Trounce IA. Mitochondrial dysfunction and complications associated with diabetes. Biochim Biophys Acta 2013; 1840: 1404-1412.
3. Ilkun O and Boudina S. Cardiac dysfunction and oxidative stress in the metabolic syndrome: an update on antioxidant therapies. Curr Pharm Des 2013; 19: 4806-4817
4. Chacko BK et al. Methods for defining distinct bioenergetic profiles in platelets, lymphocytes, mono-cytes, and neutrophils, and the oxidative burst from human blood. Lab Invest 2013; 93: 690-700.
5. Wallace DC. Bioenergetic Origins of Complexity and Disease. Cold Spring Harb. Symp Quant Biol 2011; 76: 1-16.
6. Ferrick DA, Neilson A, Beeson C. Advances in measuring cellular bioen¬ergetics using extracellular flux. Drug Discov Today 2008; 13: 268-274.
7. Nicholls DG, Darley-Usmar VM, Wu M, Jensen PB, Rogers GW, Ferrick DA. Bioenergetic profile experiment using C2C12 myoblast cells. J Vis Exp 2010; 6;(46).
8. Hill BG et al. Integration of cellular bioenergetics with mitochondrial quality control and autophagy. Biol Chem 2012; 393: 1485-1512.
9. Dranka BP, et al. Assessing bioenergetic function in response to oxidative stress by metabolic profiling. Free Radic Biol Med 2011; 51: 1621-1635.
10. Brand MD and Nicholls DG. Assessing mitochondrial dysfunction in cells. Biochem J 2011; 435: 297-312.
11. Kuznetsov AV, Veksler V, Gellerich FN, Saks V, Margreiter R, Kunz WS. Analysis of mitochondrial function in situ in permeabilized muscle fibers, tissues and cells. Nature Protocols 2008; 3(6): 965-976.
12. Duicu O, Gheorgheosu D, Mirica N, Trancotă S, Dehelean C, Firă-Mladinescu O, Muntean D. High-resolution respirometry with multiple substrates titration in permeabilized myocardial fibers. Rev Med Chir Soc Med Nat Iasi 2012; 116(1): 207-213.

Additional Files

Published

2015-06-30

Issue

Vol. 119 No. 2 (2015): The Medical-Surgical Journal

Section

BASIC SCIENCES

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