Advances in the quantification of mitochondrial function in primary human immune cells through extracellular flux analysis
Name
Nicholas-2017-Advances in the quantification o.pdf
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2.94 MB
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Adobe PDF
Checksum (MD5)
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Author(s)
Nicholas, Dequina
Raval, Forum M.
Ip, Blanche C.
Habib, Chloe
Ritou, Eleni
Grammatopoulos, Tom N.
Steenkamp, Devin
Dooms, Hans
Apovian, Caroline M.
Nikolajczyk, Barbara S.
Date Issued
February 2017
Journal
PLoS ONE
Publisher
Public Library of Science
Citation
Nicholas, Dequina; Proctor, Elizabeth A.; Raval, Forum M.; Ip, Blanche C.; Habib, Chloe; Ritou, Eleni; Grammatopoulos, Tom N et al. “Advances in the Quantification of Mitochondrial Function in Primary Human Immune Cells through Extracellular Flux Analysis.” Edited by Nades Palaniyar. PLOS ONE 12, no. 2 (February 2017): e0170975 © 2017 Nicholas et al
Version
Final published version
Abstract
Numerous studies show that mitochondrial energy generation determines the effectiveness of immune responses. Furthermore, changes in mitochondrial function may regulate lymphocyte function in inflammatory diseases like type 2 diabetes. Analysis of lymphocyte mitochondrial function has been facilitated by introduction of 96-well format extracellular flux (XF96) analyzers, but the technology remains imperfect for analysis of human lymphocytes. Limitations in XF technology include the lack of practical protocols for analysis of archived human cells, and inadequate data analysis tools that require manual quality checks. Current analysis tools for XF outcomes are also unable to automatically assess data quality and delete untenable data from the relatively high number of biological replicates needed to power complex human cell studies. The objectives of work presented herein are to test the impact of common cellular manipulations on XF outcomes, and to develop and validate a new automated tool that objectively analyzes a virtually unlimited number of samples to quantitate mitochondrial function in immune cells. We present significant improvements on previous XF analyses of primary human cells that will be absolutely essential to test the prediction that changes in immune cell mitochondrial function and fuel sources support immune dysfunction in chronic inflammatory diseases like type 2 diabetes.
MIT Department
Massachusetts Institute of Technology. Department of Biological Engineering
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Creative Commons Attribution 4.0 International License
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DOI of Published Version
https://doi.org/10.1371/journal.pone.0170975
