Calabria 2019 MiPschool Coimbra

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Elisa Calabria
Investigating mitochondrial function in blood cells by high resolution respirometry: contribution to an interlaboratory study. Mitoeagle WG4.

Link: MitoEAGLE

Calabria E, Aasander Frostner E, Castelo MP, Danila MD, Garcia-Souza LF, Gnaiger E, Keppner G, Krako Jakovljevic N, Labieniec-Watala M, Leilu T, Michalak S, Pichler I, Piel S, Rybacka-Mossakowska J, Siewiera K, Silaidos C, Sumbalova Z, Volani C, Watala C (2019)

Event: MiPschool Coimbra 2019

COST Action MitoEAGLE

The sensitivity of high resolution respirometry (HRR) has made it possible to carry out reliable measurements of oxygen consumption even in blood cells, opening up new scenarios for the mitochondrial function / dysfunction in health conditions, but also to study the pathophysiology of many diseases or the effects of effects of interventions. However, the premises of all this must be sought in the validation and comparison of the preparatory procedures, from blood collection to cell purification and in the protocols adopted for the analysis. Here are presented the procedures adopted for peripheral blood mononuclear cells (PBMC) [1,2] and for platelets (PLT) [3, 4], also in relation to other laboratories participating in WG4. Crucial points emerged and the various solutions adopted will be highlighted. We found that in the isolation of cells different approaches can be used, and the results are not always divergent.


Bioblast editor: Plangger M O2k-Network Lab: IT Verona Calabria E, SE Lund Elmer E, IT Bolzano Pichler I, RO Timisoara Muntean DM, AT Innsbruck Oroboros, AT Innsbruck Gnaiger E, DE Freising Klingenspor M, RS Belgrade Lalic NM, PL Lodz Watala C, PL Poznan Michalak S, US PA Philadelphia Kilbaugh T, DE Giessen Eckert GP, SK Bratislava Sumbalova Z


Labels: MiParea: Respiration, Instruments;methods 


Tissue;cell: Blood cells 



HRR: Oxygraph-2k, O2k-Protocol 


Affiliations

Calabria E(1), Aasander Frostner E(2), Castelo MP(3), Danila MD(4), Garcia-Souza LF(5), Gnaiger E(5), Keppner G(6), Krako Jakovljevic N(7), Labieniec-Watala M(8), Leilu T(8), Michalak S(9), Pichler I(10), Piel S(1), Rybacka-Mossakowska J(9), Siewiera K(12), Silaidos C(13), Sumbalova Z(14), Volani C(3), Watala C(8)
  1. Dept Neuroscience, Biomedicine Movement Sciences, Univ Verona, IT
  2. Mitochondrial Medicine, Lund Univ, SE
  3. Inst Biomedicine, Eurac Research, IT
  4. Victor Babes Univ Medicine Pharmacy, Timisoara, RO
  5. Oroboros Instruments, Innsbruck, AT
  6. Else Kröner Fresenius Center, Technical Univ Munich, DE
  7. Fac Medicine, Univ Belgrade, RS
  8. Dept Medical Biophysics, Univ Lodz, PL
  9. Dept Neurochemistry Neuropathology, Poznan Univ Medical Sciences, PL
  10. Center Biomedicine, European Academy of Bolzano, IT
  11. Anesthesia Critical Care, The Children’s Hospital Philadelphia, US
  12. Dept Haemostasis Haemostatic Disorders, Medical Univ Lodz, PL
  13. Dept Nutrition Prevention & Therapy, Justus Liebig Univ, Gießen DE
  14. Fac Medicine, Comenius Univ, SK. - elisa.calabria@univr.it

References

  1. Michalak S, Florczak-Wyspiańska J, Rybacka-Mossakowska J, Ambrosius W, Osztynowicz K, Baszczuk A, Kozubski W, Wysocka E (2017) Mitochondrial respiration in intact peripheral blood mononuclear cells and Sirtuin 3 activity in patients with movement disorders. Oxid Med Cell Longev 2017:9703574.
  2. Sumbalova Z, Droescher S, Hiller E, Chang SC, Garcia-Souza LF, Calabria E, Volani C, Krumschnabel G, Gnaiger E (2018) O2k-Protocols: Isolation of blood cells for HRR. Mitochondr Physiol Network 17:1-16.
  3. Sjövall F, Ehinger JK, Marelsson SE, Morota S, Frostner EA, Uchino H, Lundgren J, Arnbjörnsson E, Hansson MJ, Fellman V, Elmér E (2013) Mitochondrial respiration in human viable platelets--methodology and influence of gender, age and storage. Mitochondrion 13:7-14.
  4. Gatterer H, Menz V, Salazar-Martinez E, Sumbalova Z, Garcia-Souza LF, Velika B, Gnaiger E, Burtscher M (2018) Exercise performance, muscle oxygen extraction and blood cell mitochondrial respiration after repeated-sprint and sprint interval training in hypoxia: A pilot study. J Sports Sci Med 17:339-47.