Hoppel 2015 Abstract MiPschool London 2015
|Human blood cell types as study models of mitochondrial respiration and H2O2 production.|
Event: MiPschool London 2015
Respirometric and fluorometric measurements are performed by the use of living cells, isolated mitochondria or tissue homogenate for many research aims, whereas permeabilized fibres obtained from muscle biopsies are the gold standard in research of specific questions in sport science. Collecting muscle biopsies is associated with numerous ethical, sanitary and organisational difficulties and thus a simplified and less invasive method of obtaining samples from human tissues for research on questions in health and disease is highly in need and would definitely boost the acquisition of knowledge on mitochondrial functions in humans in various research fields. Therefore, human blood cells, such as thrombocytes (synonym: platelets), lymphocytes and monocytes (synonym: Peripheral Blood Mononuclear Cell) and granulocytes (synonym: polymorphonuclear leukocytes) would seem to represent an attractive alternative, given the relative ease and limited invasiveness with which blood samples can be gained. However, although human platelets have been suggested as a model for research of mitochondrial metabolism , only few respirometric studies using human blood cells were published so far. Furthermore, a comparison of the different human blood cell types as those mentioned above is also missing. The simultaneous measurement of mitochondrial O2 consumption and H2O2 production by use of the OROBOROS Oxygraph-2k and the Fluorescence-module allows the assessment in detail of both mitochondrial energetics and the associated formation of ROS using relatively small samples. This method would thus seem ideally suited to study the energetics of various blood cell types .
A further important major advantage of the use of human blood cells over the use of muscle biopsies would be the possibility to collect and store samples for later use by freezing them. While tissue samples have to be analysed as soon as possible after they have been obtained, intact cells afford the opportunity of long time storage by cyropreservation using a freezing medium prepared by mixing dimethyl sulfoxide and fetal bovine serum at a specific ratio . Given that there is still an enormous lack of knowledge regarding the possible influence of freezing and the frozen preservation on respiration of intact and permeabilized cells, further research on this topic is definitely required.
In consequence, in addition to standardization of cell separation protocols and of experimental media, research on the stability of biological samples related to cyropreservation has to be performed and documented in Standard Operating Procedures to ensure optimised quality in all these regards for future research using human blood cells.
Labels: MiParea: Instruments;methods, Exercise physiology;nutrition;life style
Stress:Cryopreservation Organism: Human Tissue;cell: Blood cells Preparation: Intact cells
HRR: Oxygraph-2k, O2k-Fluorometer
1-Dept Sport Sc, Univ Innsbruck
2-D. Swarovski Research Lab, Dept Visceral, Transplant Thoracic Surgery, Medical Univ Innsbruck
3-Oroboros Instruments; Innsbruck, Austria – [email protected]
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