Cellular stress is central to the understanding of pathological mechanisms and the development
of new therapeutic strategies and serves as a biomarker for disease progression in neurodegeneration,
diabetes, cancer, cardiovascular and other chronic diseases. The common cellular
stress assay (CSA) based on Seahorse technology in peripheral blood mononuclear cells (PBMCs)
shows inconsistent results, which prevents its use as a biomarker for the progression of chronic
diseases. Therefore, the aim of this study was to investigate potential factors that affect the CSA in
PBMCs. We measured the CSA parameters in PBMCs from study participants and compared the
results according to the potential factors, namely, the PBMC isolation method, age, seasonal variation
and the gender of the study participants. PBMCs were isolated by OptiPrep® and RobosepTM-S
methods. PBMCs isolated with the OptiPrep method showed much higher extracellular acidification
and higher respiration compared to Robosep-isolated cells. Moreover, OptiPrep-isolated cells showed
a higher number of outliers for the proton production rate (PPR) and a high respiratory quotient,
indicating impurities with other cells, such as platelets, and technical inconsistencies. PBMCs from
older individuals showed higher maximal respiration, spare capacity and extracellular acidification
than younger participants. Additionally, in winter, maximal respiration and spare capacity decreased.
From spring until early autumn, spare capacity and maximal respiration continuously increased.
Elderly males also showed higher basal respiration, spare capacity and extracellular acidification
than females. In conclusion, the findings of this study clearly demonstrate that the results of CSA
parameters measured in PBMCs are influenced by the PBMC isolation method, age, seasonal variation
and gender. Therefore, we recommend that researchers and physicians properly interpret the
results of CSA parameters in PBMCs by considering these factors. It is important to use separate
CSA evaluation standards based on the isolation method, age, gender and season-dependent factors.
To assess the cellular stress situation in PBMCs, both extracellular acidification and mitochondrial
respiration should be taken into account. Further study of additional factors, such as mitochondrial
mass, should be conducted to improve the measurement of CSA parameters for the assessment of the
real mitochondrial fitness.
| Identifer | oai:union.ndltd.org:DRESDEN/oai:qucosa:de:qucosa:91340 |
| Date | 13 May 2024 |
| Creators | Tessema, Belay, Riemer, Janine, Sack, Ulrich, König, Brigitte |
| Publisher | MDPI |
| Source Sets | Hochschulschriftenserver (HSSS) der SLUB Dresden |
| Language | English |
| Detected Language | English |
| Type | info:eu-repo/semantics/publishedVersion, doc-type:article, info:eu-repo/semantics/article, doc-type:Text |
| Rights | info:eu-repo/semantics/openAccess |
| Relation | 13118 |
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