Abstract: Sumbalova Z, Droescher S, Hiller E, Chang SC, Garcia-Souza LF, Calabria E, Volani C, Krumschnabel G, Gnaiger E (2018) O2k-Protocols: Isolation of peripheral blood mononuclear cells and platelets from human blood for HRR. Mitochondr Physiol Network 21.17(03):1-16.
Respiratory assessment of human health and disease can be performed with isolated mitochondria, tissue homogenate or permeabilized fibers prepared from tissue biopsies. The collection of tissue biopsies is invasive, limiting its applicability for large-scale screening. An alternative is the use of blood cells, which can be obtained in a far less invasive sampling procedure. Blood cells obtained by venipuncture are usually separated to obtain 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. A promising field of innovation is the extended storage or even cryopreservation of blood cells for later use in respirometry. In the present study, isolation methods are described to obtain blood cells for high-resolution respirometry (HRR), methodological details of the isolation procedure are highlighted, and the protocols for the respiratory characterization of platelets and PBMCs are presented.
• O2k-Network Lab: AT_Innsbruck_OROBOROS
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, ROX HRR: Oxygraph-2k, O2k-Protocol