Talk:MitoEAGLE blood cells 1

Phase 1: Innsbruck retreat Jan 2018

Aims

• Prepare MitoEAGLE Poznan 2018
• Complete Version 01 of joint publication
• Discuss and update MitoEAGLE preprint 2017-09-21

Participants

• Chang SC, Duicu OM, Garcia-Souza LF, Gnaiger E, Labieniec-Watala M, Michalak S, Siewiera K, Silaidos C, Sumbalova Z, Volani C

Agenda

2018-01-29

• Start-up discussion
• Manuscript structure
• Title
• Abstract
• Publication website

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Abstract Version beta 01

1. The evaluation of mitochondrial function remains crucial for the diagnosis of a spectrum of human pathologies. Peripheral blood provides an easily accessible source of biological material like cells. However, mitochondrial respiratory studies in blood cells still require standardization. The procedures applied in bioenergetic assessments involve multiple steps including pre-analytical, analytical and normalization phase. Therefore, the aim of this report is to review and summarize methods using peripheral blood mononuclear cells and platelets for mitochondrial respiratory studies. For this purpose we have used original data from multiple laboratories combined with literature data.
2. This methodological review is not primarily to present results on patients, but to characterize control groups for pathophysiological studies. During pre-analytical phase smoking, alcohol intake, BMI threshold, lifestyle intervention and medications should be considered for inclusion / exclusion of controls. Moreover, all subjects should be matched for sex, age and ethnicity. Conditions like time of sampling, fasting and resting state are terms of standardization.
3. Anticoagulants are used for whole blood sampling /e.g. K-EDTA, lithium heparin, sodium citrate/ and discussed for their effect on cells isolation, yield, cells fraction purity and respiration.
4. The time and temperature during transport and storage of whole blood before specific cells separation influences next analytical phases and affect final outcome. Characterization of whole blood provides the basis for inclusion / exclusion for further processing of the sample.
5. Separation procedures depend on the type of cells required for respirometric studies / PLT, PBMC or PLT and PBMC/. Media used, centrifugation conditions, cell counting and cell viability methods are under discussion. Purity of preparation / e.g. PLT contamination in PBMCs fraction, purity of PLT fraction/, recovery and yield of cells from the whole blood are also presented in this report.
6. There are limited data 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 could significantly affect the measurements.