ArticleFrontiers in immunology2025
Comprehensive real-time metabolic profiling of peripheral blood mononuclear cells reveals important methodological considerations for immunometabolism research.
Article in Frontiers in immunology, 2025. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 6 papers.
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Who cites it
6 citing papers in PubMed.
- Peripheral Mononuclear Cells Metabolomics in Obstructive Sleep Apnea Syndrome (OSAS): An Exploratory Pilot Study of Immunometabolic Signatures.International journal of molecular sciences · 2026Article
- Autofluorescence lifetime imaging resolves cell heterogeneity within peripheral blood mononuclear cells.Biophotonics discovery · 2026Article
- A Case Report of Reversible Mitochondrial Bioenergetic Dysfunction in PBMCs in Anti-GAD65-Associated Cerebellar Ataxia.Cerebellum (London, England) · 2026Article
- Autofluorescence lifetime imaging resolves cell heterogeneity within peripheral blood mononuclear cells.bioRxiv : the preprint server for biology · 2026Article
- Orchestrating the gut microbiota-mitochondrial-immune axis in gynecological diseases: mechanisms and dual-targeting therapeutic strategies.Frontiers in reproductive health · 2026Review
- Mitochondrial respiration in human peripheral blood mononuclear cells: methodology and influence of permeabilization and storage.Frontiers in molecular biosciences · 2026Article
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Authors and funding
4 authors.
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Abstract
Background/Introduction: The utility of measuring real-time cellular bioenergetics of peripheral blood mononuclear cells (PBMCs) as biomarkers in disease monitoring, such as the bioenergetic health index, is of emerging interest. However, various experimental factors can impact the accuracy and reproducibility of these measurements.Methods: PBMC bioenergetics were probed in real-time using extracellular flux analysis to identify optimal seeding density and injection protocol. Using a modified protocol, we assessed the extent to which blood processing time and isolation method (SepMate™ vs. EasySep™ Direct) influence PBMC bioenergetics under basal and stimulated conditions. Advanced metabolic control analysis including mitochondrial and glycolytic ATP supply flux, respiratory control ratio, bioenergetic health index, and mitochondrial toxicity index were used to identify and quantify PBMC bioenergetics. Results: Measures of metabolic profiling such as mitochondrial respiration, glycolytic activity, ATP supply flux, and respiratory control ratio were significantly diminished in PBMCs due to blood processing delay (48-72 hours) and were influenced by isolation method. Extended blood processing time significantly lowered T cell activation capacity in PBMCs, evidenced by decreased responses of mitochondrial and glycolytic ATP supply to CD3/CD28 activation. Discussion/Conclusion: This study demonstrates that key experimental variables including blood processing time and isolation method critically affect the reliability and biological relevance of PBMC metabolic assessments, highlighting the importance of protocol standardisation for accurate bioenergetic biomarker measurements.
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