Measurement of mitochondrial respiration in adherent cells by Seahorse XF96 Cell Mito Stress Test

Xia Gu; Yuan Ma; Yi Liu; Qiang Wan

doi:10.1016/j.xpro.2020.100245

Measurement of mitochondrial respiration in adherent cells by Seahorse XF96 Cell Mito Stress Test

STAR Protoc. 2020 Dec 30;2(1):100245. doi: 10.1016/j.xpro.2020.100245. eCollection 2021 Mar 19.

Authors

Xia Gu¹, Yuan Ma², Yi Liu³, Qiang Wan⁴

Affiliations

¹ School of Medicine, Cheeloo College of Medicine, Shandong University, Jinan, Shandong 250012, China.
² Renal Division, Peking University First Hospital, Peking University Institute of Nephrology, Beijing 100034, China.
³ Department of Pulmonary and Critical Care Medicine, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong 250012, China.
⁴ Department of Endocrinology, Shandong Provincial Hospital, Cheeloo College of Medicine, Shandong University, Jinan, Shandong 250012, China.

Abstract

Mitochondria play pivotal roles in cellular energy metabolism. Most of the intracellular adenosine triphosphate (ATP) is generated by mitochondrial respiration. The Cell Mito Stress Test is a common method to measure the key parameters of mitochondrial respiration. Here, we use the human cell line HK-2 as an example to present the procedures to quantify the oxygen consumption rate using a Seahorse XFe96 extracellular flux analyzer. For complete details on the use and execution of this protocol, please refer to Ma et al. (2020).

Keywords: Cell Biology; Cell culture; Cell-based Assays; Metabolism; Molecular Biology.

Publication types

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

MeSH terms

Cell Adhesion
Cell Line
Humans
Mitochondria / metabolism*
Oxygen Consumption*