ArticleJournal of visualized experiments : JoVE2016
An Optimized Protocol to Analyze Glycolysis and Mitochondrial Respiration in Lymphocytes.
Article in Journal of visualized experiments : JoVE, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 29 papers.
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
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Who cites it
29 citing papers in PubMed, 38 citations in OpenAlex.
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- Transcriptional profiling of lung macrophages following ozone exposure in mice identifies signaling pathways regulating immunometabolic activation.Toxicological sciences : an official journal of the Society of Toxicology · 2024Article
- GABA-mediated inhibition of human CD4EBioMedicine · 2024Article
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- Excess Dietary Sugar Alters Colonocyte Metabolism and Impairs the Proliferative Response to Damage.Cellular and molecular gastroenterology and hepatology · 2023Article
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- Using Real-Time Cell Metabolic Flux Analyzer to Monitor Osteoblast Bioenergetics.Journal of visualized experiments : JoVE · 2022Article
- Mitochondrial metabolism determines the functional status of human sperm and correlates with semen parameters.Frontiers in cell and developmental biology · 2022Article
- Glycolysis, monocarboxylate transport, and purinergic signaling are key events inFrontiers in immunology · 2022Article
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- Novel aspects of regulatory T cell dysfunction as a therapeutic target in giant cell arteritis.Annals of the rheumatic diseases · 2022Article
- Assessment of Cellular Bioenergetics in Mouse Hematopoietic Stem and Primitive Progenitor Cells using the Extracellular Flux Analyzer.Journal of visualized experiments : JoVE · 2021Article
- Utilization of Human Samples for Assessment of Mitochondrial Bioenergetics: Gold Standards, Limitations, and Future Perspectives.Life (Basel, Switzerland) · 2021Review
- Article
- Mitochondrial and Glycolytic Capacity of Peripheral Blood Mononuclear Cells Isolated From Diverse Poultry Genetic Lines: Optimization and Assessment.Frontiers in veterinary science · 2021Article
- Capacitation increases glucose consumption in murine sperm.Molecular reproduction and development · 2020Article
- Analysis of Human Natural Killer Cell Metabolism.Journal of visualized experiments : JoVE · 2020Article
- High-content fluorescence imaging with the metabolic flux assay reveals insights into mitochondrial properties and functions.Communications biology · 2020Article
Corrections and comments
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
Abstract
Lymphocytes respond to a variety of stimuli by activating intracellular signaling pathways, which in turn leads to rapid cellular proliferation, migration and differentiation, and cytokine production. All of these events are tightly linked to the energy status of the cell, and therefore studying the energy-producing pathways may give clues about the overall functionality of these cells. The extracellular flux analyzer is a commonly used device for evaluating the performance of glycolysis and mitochondrial respiration in many cell types. This system has been used to study immune cells in a few published reports, yet a comprehensive protocol optimized particularly for lymphocytes is lacking. Lymphocytes are fragile cells that survive poorly in ex vivo conditions. Oftentimes lymphocyte subsets are rare, and working with low cell numbers is inevitable. Thus, an experimental strategy that addresses these difficulties is required. Here, we provide a protocol that allows for rapid isolation of viable lymphocytes from lymphoid tissues, and for the analysis of their metabolic states in the extracellular flux analyzer. Furthermore, we provide results of experiments in which the metabolic activities of several lymphocyte subtypes at different cell densities were compared. These observations suggest that our protocol can be used to achieve consistent, well-standardized results even at low cell concentrations, and thus it may have broad applications in future studies focusing on the characterization of metabolic events in immune cells.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.