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MitoEAGLE blood cells 1
Additional label MitoFitPublication  + , MitoEAGLEPublication  +
Coupling states LEAK  + , ROUTINE  + , OXPHOS  + , ET  +
Diseases Aging;senescence  +
Enzyme Marker enzyme  +
Has abstract [[File:Workflow_MitoEAGLE blood cells.png|
[[File:Workflow_MitoEAGLE blood cells.png|500px|right|thumb|Fig. 1. Workflow and topics. SUIT protocols for measurement of respiration in intact cells (viable cells, vce) and permeabilized cells (pce).]] ::: '''Version beta 01''' :::::# The evaluation of mitochondrial function remains crucial for the diagnosis of a spectrum of human pathologies. Peripheral blood provides an easily accessible source of primary human cells. However, mitochondrial respiratory studies in blood cells still require standardization. The procedures applied in bioenergetic assessments involve multiple steps in the pre-analytical, analytical and data analysis phase. The aim of this review is to summarize methods of preparing and applying peripheral blood mononuclear cells (PBMC) and platelets (PLT) for mitochondrial respiratory studies. For this purpose we report original data from several laboratories and literature data. :::::# We do not present results on patients, but characterize healthy control groups. Exclusion criteria in the pre-analytical phase are related to smoking, alcohol intake, BMI, lifestyle interventions, and medications. In pathophysiological studies, subjects have to be matched with controls for sex, age and genetic background. Time of sampling, fasting and resting state require standardization. :::::# Anticoagulants are used for whole blood sampling (''e.g.'', K-EDTA, lithium heparin, sodium citrate). Protocols are optimized on the basis of yield, cell fraction purity and respiratory function. :::::# The time and temperature during transport and storage of whole blood has to be monitored carefully and evaluated in terms of final outcome. Characterization of whole blood provides rigorous exclusion criteria for controls. :::::# Separation procedures depend on the type of cells harvested for respirometric studies (PLT, PBMC, or both). Media, centrifugation conditions, cell counting and cell viability methods are evaluated. Results on purity of preparations (''e.g.'', PLT contamination in PBMC fraction, purity of PLT fraction), recovery and yield of cells are compared in studies using a variety of cell separation methods. :::::# Information is lacking on the effect of storage and conditions during processing of isolated cell types before respirometry. Temperature, media, antibiotics, proteinase inhibitors cocktail, density of cells during storage, tilting of cell fraction might significantly affect the measurements. :::::# Specific protocols are used for respiration assessments in intact and permeabilized. The type of medium used for respirometry (MiR05, RPMI, plasma) needs further evaluation. :::::# Normalization is important to interpret and compare the results of respiratory studies. It should include data on contamination of PBMC with PLT, cell viability (''e.g.'', succinate test), flux control ratios, total protein concentration, citrate synthase activity. Some other markers, like clusters of differentiation specific for leukocytes and platelets can be taken into consideration. :::::# Finally, we summarize emerging recommendations for mitochondrial respiratory studies in intact and permeabilized PBMC and PLT towards building a data base of healthy subjects that can be used as controls in clinical studies.
n be used as controls in clinical studies.  +
Has info [[WG4_MitoEAGLE_data:_blood_and_cultured_cells#TG4.1_Blood |MitoEAGLE blood cells group (WG 4)]]  +
Has title MitoEAGLE blood cells group (2018) An interlaboratory guide through procedures for mitochondrial respiratory studies with intact and permeabilized peripheral blood mononuclear cells and platelets. - Updated: 2018-02-23  +
Mammal and model Human  +
MiP area Respiration  + , Instruments;methods  + , mtDNA;mt-genetics  + , nDNA;cell genetics  + , Gender  +
Pathways F  + , N  + , S  + , Gp  + , CIV  + , NS  + , Other combinations  + , ROX  +
Preparation Intact cells  + , Permeabilized cells  +
Tissue and cell Blood cells  + , Platelet  +
Was published in journal MitoEAGLE +
Was published in year 2018  +
Was written by Calabria E + , Duicu OM + , Garcia-Souza LF + , Gnaiger E + , Labieniec-Watala M + , Michalak S + , Siewiera K + , Silaidos C + , Sumbalova Z + , Volani C + , Aasander Frostner E + , Chamkha I + , Danila MD + , Keppner G + , Lelcu T + , Pavlovic K + , Kidere D + , Piel S + , Rybacka-Mossakowska J + , Swiniuch D + , Vujacic-Mirski K + , Watala C + , Chang SC +
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MitoEAGLE blood cells 1  +
Categories MitoGlobal , Publications
Modification date
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08:29:09, 5 August 2019  +
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