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Property "Has-20abstract" (as page type) with input value "-5B-5BFile-2D3AWorkflow_MitoEAGLE-20blood-20cells.png-7C500px-7Cright-7Cthumb-7CFig.-201.-20Workflow-20and-20topics.-20SUIT-20protocols-20for-20measurement-20of-20respiration-20in-20living-20cells-20(viable-20cells,-20vce-3B-20dead-20cells,-20dce)-20and-20permeabilized-20cells-20(pce).-5D-5D-0A-2D3A-2D3A-2D3A-20-27-27'Version-20beta-2001-27-27'-0A-2D3A-2D3A-2D3A-2D3A-2D3A-23-20The-20evaluation-20of-20mitochondrial-20function-20remains-20crucial-20for-20the-20diagnosis-20of-20a-20spectrum-20of-20human-20pathologies.-20Peripheral-20blood-20provides-20an-20easily-20accessible-20source-20of-20primary-20human-20cells.-20However,-20mitochondrial-20respiratory-20studies-20in-20blood-20cells-20still-20require-20standardization.-20The-20procedures-20applied-20in-20bioenergetic-20assessments-20involve-20multiple-20steps-20in-20the-20pre-2Danalytical,-20analytical-20and-20data-20analysis-20phase.-20The-20aim-20of-20this-20review-20is-20to-20summarize-20methods-20of-20preparing-20and-20applying-20peripheral-20blood-20mononuclear-20cells-20(PBMC)-20and-20platelets-20(PLT)-20for-20mitochondrial-20respiratory-20studies.-20For-20this-20purpose-20we-20report-20original-20data-20from-20several-20laboratories-20and-20literature-20data.-20-0A-2D3A-2D3A-2D3A-2D3A-2D3A-23-20We-20do-20not-20present-20results-20on-20patients,-20but-20characterize-20healthy-20control-20groups.-20Exclusion-20criteria-20in-20the-20pre-2Danalytical-20phase-20are-20related-20to-20smoking,-20alcohol-20intake,-20BMI,-20lifestyle-20interventions,-20and-20medications.-20In-20pathophysiological-20studies,-20subjects-20have-20to-20be-20matched-20with-20controls-20for-20sex,-20age-20and-20genetic-20background.-20Time-20of-20sampling,-20fasting-20and-20resting-20state-20require-20standardization.-0A-2D3A-2D3A-2D3A-2D3A-2D3A-23-20Anticoagulants-20are-20used-20for-20whole-20blood-20sampling-20(-27-27e.g.-27-27,-20K-2DEDTA,-20lithium-20heparin,-20sodium-20citrate).-20Protocols-20are-20optimized-20on-20the-20basis-20of-20yield,-20cell-20fraction-20purity-20and-20respiratory-20function.-20-0A-2D3A-2D3A-2D3A-2D3A-2D3A-23-20The-20time-20and-20temperature-20during-20transport-20and-20storage-20of-20whole-20blood-20has-20to-20be-20monitored-20carefully-20and-20evaluated-20in-20terms-20of-20final-20outcome.-20Characterization-20of-20whole-20blood-20provides-20rigorous-20exclusion-20criteria-20for-20controls.-0A-2D3A-2D3A-2D3A-2D3A-2D3A-23-20Separation-20procedures-20depend-20on-20the-20type-20of-20cells-20harvested-20for-20respirometric-20studies-20(PLT,-20PBMC,-20or-20both).-20Media,-20centrifugation-20conditions,-20cell-20counting-20and-20cell-20viability-20methods-20are-20evaluated.-20Results-20on-20purity-20of-20preparations-20(-27-27e.g.-27-27,-20PLT-20contamination-20in-20PBMC-20fraction,-20purity-20of-20PLT-20fraction),-20recovery-20and-20yield-20of-20cells-20are-20compared-20in-20studies-20using-20a-20variety-20of-20cell-20separation-20methods.-0A-2D3A-2D3A-2D3A-2D3A-2D3A-23-20Information-20is-20lacking-20on-20the-20effect-20of-20storage-20and-20conditions-20during-20processing-20of-20isolated-20cell-20types-20before-20respirometry.-20Temperature,-20media,-20antibiotics,-20proteinase-20inhibitors-20cocktail,-20density-20of-20cells-20during-20storage,-20tilting-20of-20cell-20fraction-20might-20significantly-20affect-20the-20measurements.-0A-2D3A-2D3A-2D3A-2D3A-2D3A-23-20Specific-20protocols-20are-20used-20for-20respiration-20assessments-20in-20living-20and-20permeabilized.-20The-20type-20of-20medium-20used-20for-20respirometry-20(MiR05,-20RPMI,-20plasma)-20needs-20further-20evaluation.-0A-2D3A-2D3A-2D3A-2D3A-2D3A-23-20Normalization-20is-20important-20to-20interpret-20and-20compare-20the-20results-20of-20respiratory-20studies.-20It-20should-20include-20data-20on-20contamination-20of-20PBMC-20with-20PLT,-20cell-20viability-20(-27-27e.g.-27-27,-20succinate-20test),-20flux-20control-20ratios,-20total-20protein-20concentration,-20citrate-20synthase-20activity.-20Some-20other-20markers,-20like-20clusters-20of-20differentiation-20specific-20for-20leukocytes-20and-20platelets-20can-20be-20taken-20into-20consideration.-0A-2D3A-2D3A-2D3A-2D3A-2D3A-23-20Finally,-20we-20summarize-20emerging-20recommendations-20for-20mitochondrial-20respiratory-20studies-20in-20living-20and-20permeabilized-20PBMC-20and-20PLT-20towards-20building-20a-20data-20base-20of-20healthy-20subjects-20that-20can-20be-20used-20as-20controls-20in-20clinical-20studies." contains invalid characters or is incomplete and therefore can cause unexpected results during a query or annotation process.

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