Sumbalova 2016b Abstract MitoFit Science Camp 2016

From Bioblast
Human blood cells: isolation and HRR.

Link:

Sumbalova Z, Droescher S, Hiller E, Chang S, Garcia-Souza Luiz F, Calabria E, Volani C, Krumschnabel G, Gnaiger E (2016)

Event: MitoFit Science Camp 2016 Kuehtai AT

Respiratory assessment of human health and disease is often performed with isolated mitochondria, tissue homogenate or permeabilized fibres prepared from tissue biopsies. However, the collection of tissue biopsies is invasive and experimentally cumbersome, limiting its applicability. An alternative is the use of blood cells, which can be obtained in a far less invasive sampling procedure and can be at least temporarily stored after collection for later use in respirometric measurements. Blood cells obtained by venipuncture are then usually separated to obtain either platelets (PLT) or a mixed population of immune cells subsumed as peripheral blood mononuclear cell (PBMCs), both of which have been successfully applied to characterize respiratory phenotypes of human diseases.

In the present study, isolation methods to obtain blood cells for high-resolution respirometry (HRR) are described, important aspects of the isolation procedure are highlighted, and the protocols for the respiratory characterization of platelets and PBMCs are presented.

β€’ Keywords: PBMC

β€’ O2k-Network Lab: AT Innsbruck Oroboros, IT Verona Calabria E


Affiliations

1-Pharmacobiochem Lab, Fac Med, Comenius Univ, Bratislava, Slovakia; 2-Oroboros Instruments; 3-Dept Visceral, Transplant Thoracic Surgery, D. Swarovski Research Lab, Medical Univ Innsbruck, Austria. – [email protected]

References

  1. Chacko BK, Kramer PA, Ravi S, Johnson MS, Hardy RW, Ballingern SW, Darley-Usmar VM (2013) Methods for defining distinct bioenergetic profiles in platelets, lymphocytes, monocytes, and neutrophils, and the oxidative burst from human blood. Lab Invest 93:690–700.
  2. Karabatsiakis A, BΓΆck C, Salinas-Manrique J, Kolassa S, Calzia E, Dietrich DE, Kolassa IT (2014) Mitochondrial respiration in peripheral blood mononuclear cells correlates with depressive subsymptoms and severity of major depression. Transl Psychiatry 4:e397.
  3. SjΓΆvall F, Ehinger JK, Marelsson SE, Morota S, Frostner EA, Uchino H, Lundgren J, ArnbjΓΆrnsson E, Hansson MJ, Fellman V, ElmΓ©r E (2013) Mitochondrial respiration in human viable platelets-methodology and influence of gender, age and storage. Mitochondrion 13:7-14.
  4. Harmonized SUIT protocols


Labels: MiParea: Respiration, Instruments;methods 


Organism: Human  Tissue;cell: Blood cells, Lymphocyte, Platelet  Preparation: Intact cells, Permeabilized cells 


Coupling state: LEAK, ROUTINE, OXPHOS, ET  Pathway: F, N, S, Gp, CIV, NS, Other combinations, ROX  HRR: Oxygraph-2k  Event: B1  MitoFit Science Camp 2016 

Cookies help us deliver our services. By using our services, you agree to our use of cookies.