Cizmarova 2017 MITOEAGLE Obergurgl
Event: MitoEAGLE Obergurgl 2017
Cryopreservation appears to be a beneficial and promising method for preservation of mitochondrial functions of peripheral blood mononuclear cells (PBMC) and platelets (PLT). Cryopreservation in general have significant effect on the cell recovery and viability.
In our study, we measured respiration of intact cells at 37°C using coupling control protocol extended for respirometric determination of cell viability (coupling control and cell viability protocol - CCVP). Freshly isolated or cryopreserved blood cells (3 mill of PBMC; 200 mill of PLT) were added into the 2-ml chambers of the O2k-FluoRespirometer (Oroboros Instruments, Innsbruck, Austria) containing mitochondrial respiration medium MiR06-Kit (Oroboros, AT).
The viability of PBMC determined by CCVP (in average 93.9% for fresh and 79.6% for cryopreserved PBMC) corresponded well to viability determined by Luna™ automated cell counter (96.2% and 85.4% correspondingly for fresh and cryopreserved PBMC). CCVP can be successfully used for determination of viability in platelets, that cannot be obtained by viability test in cell counter.
Standardization of methods for blood cells isolation, cryopreservation, thawing, as well as storage condition for long-term storage is necessary for quality control and data comparison. Cryopreserved PBMC as well as PLT offer a promising model for mitochondrial respiratory diagnostic studies.
Labels: MiParea: Respiration
Tissue;cell: Blood cells, Platelet Preparation: Intact cells
(1,2)Cizmarova B, (1,3)Garcia-Souza LF, (3)Menz V, (3)Burtscher M, (4)Karabatsiakis A, (1,5)Sumbalova Z, (1,6)Gnaiger E
- Daniel Swarovski Research Lab, Dept Visceral, Transplant Thoracic Surgery, Medical Univ Innsbruck, Austria
- Dept Medical Clinical Biochem, Fac Medicine, Pavol Jozef Šafárik Univ Košice, Slovakia
- Inst Sport Science, Univ Innsbruck, Austria
- Univ Ulm, Clinical Biol Psychol, Germany
- Pharmacobiochemical Lab, 3rd Dept Internal Medicine, Fac Medicine, Comenius Univ, Bratislava, Slovakia
- Oroboros Instruments, Innsbruck, Austria. - [email protected]