MiPNet21.17 BloodCellsIsolation
O2k-Protocols: Isolation of blood cells for HRR. |
OROBOROS (2016) Mitochondr Physiol Network
Abstract: Sumbalova Z, Droescher S, Hiller E, Chang S, Garcia L, Calabria E, Volani C, Krumschnabel G, Gnaiger E (2016) Isolation of blood cells for HRR. Mitochondr Physiol Network 21.17(01):1-15.
Respiratory is often performed with isolated mitochondria, tissue homogenate or permeabilized fibres prepared from tissue biopsies. Particularly the latter is invasive and experimentally cumbersome, limiting its applicability. Thus, the alternative use of blood cells, easily obtained by venipuncture, has become increasingly popular in respirometry. Typically whole blood is then 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 this protocol, 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’ O2k-Network Lab: AT_Innsbruck_OROBOROS
Labels: MiParea: Respiration, Instruments;methods
Organism: Mouse
Tissue;cell: Blood cells
Preparation: Intact cells, Permeabilized cells
Coupling state: LEAK, ROUTINE, OXPHOS, ETS"ETS" is not in the list (LEAK, ROUTINE, OXPHOS, ET) of allowed values for the "Coupling states" property.
HRR: Oxygraph-2k, O2k-Protocol
O2k-Demo, Mt-Preparation