Sumbalova 2016b Abstract MitoFit Science Camp 2016
|Human blood cells: isolation and HRR.|
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.
• Keywords: PBMC
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
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]
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- Harmonized SUIT protocols