Metabolic analysis of mouse bone-marrow-derived dendritic cells using an extracellular flux analyzer

Kazuhito Gotoh; Yurie Takata; Yuya Nakashima; Soichi Mizuguchi; Keishi Komori; Dongchon Kang

doi:10.1016/j.xpro.2021.100401

Metabolic analysis of mouse bone-marrow-derived dendritic cells using an extracellular flux analyzer

STAR Protoc. 2021 Mar 26;2(2):100401. doi: 10.1016/j.xpro.2021.100401. eCollection 2021 Jun 18.

Authors

Kazuhito Gotoh¹, Yurie Takata¹, Yuya Nakashima¹, Soichi Mizuguchi¹, Keishi Komori¹, Dongchon Kang¹

Affiliation

¹ Department of Clinical Chemistry and Laboratory Medicine, Graduate School of Medical Sciences, Kyushu University, 3-1-1, Maidashi, Higashi-ku, Fukuoka 812-8582, Japan.

Abstract

Dendritic cell (DC) maturation induced by Toll-like receptor (TLR) agonists requires the activation of downstream metabolic changes. Here, we provide a detailed protocol to measure glycolysis, mitochondrial respiration, and fatty acid oxidation in mouse bone-marrow-derived DCs with the Seahorse XF24 extracellular flux (XF) analyzer. XF analysis with the Seahorse bioanalyzer has become a standard method to measure bioenergetic functions in cells, and this protocol can be adapted to other immune cells. For complete information on using this protocol, please refer to Gotoh et al. (2018).

Keywords: Cell biology; Cell culture; Cell isolation; Cell-based assays; Immunology; Metabolism.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Animals
Bone Marrow Cells / metabolism*
Cell Culture Techniques
Cells, Cultured
Dendritic Cells / metabolism*
Fatty Acids / metabolism
Glycolysis / physiology
Male
Metabolic Flux Analysis / methods*
Mice
Mitochondria / metabolism

Substances

Fatty Acids