Robust intrinsic differences in mitochondrial respiration and H2O2 emission between L6 and C2C12 cells

Rat L6 and mouse C2C12 cell lines are commonly used to investigate myocellular metabolism. Mitochondrial characteristics of these cell lines remain poorly understood despite mitochondria being implicated in the development of various metabolic diseases. To address this need, we performed high-resolu...

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Bibliographic Details
Published in:American Journal of Physiology: Cell Physiology 2019-08, Vol.317 (2), p.C339-C347
Main Authors: Robinson, Matthew M, Sather, Bergen K, Burney, Emily R, Ehrlicher, Sarah E, Stierwalt, Harrison D, Franco, Maria Clara, Newsom, Sean A
Format: Article
Language:English
Subjects:
Carnitine
Cell lines
Electron transfer
Electron transport
Hydrogen peroxide
L-Carnitine
Metabolic disorders
Metabolism
Mitochondria
Myoblasts
Myotubes
Oxidation
Oxidative phosphorylation
Oxygen consumption
Phosphorylation
Respiration
Titration
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Summary:Rat L6 and mouse C2C12 cell lines are commonly used to investigate myocellular metabolism. Mitochondrial characteristics of these cell lines remain poorly understood despite mitochondria being implicated in the development of various metabolic diseases. To address this need, we performed high-resolution respirometry to determine rates of oxygen consumption and H2O2 emission in suspended myoblasts during multiple substrate-uncoupler-inhibitor titration protocols. The capacity for oxidative phosphorylation supported by glutamate and malate, with and without succinate, or supported by palmitoyl-l-carnitine was lower in L6 compared with C2C12 myoblasts (all P < 0.01 for L6 vs. C2C12). Conversely, H2O2 emission during oxidative phosphorylation was greater in L6 than C2C12 myoblasts (P < 0.01 for L6 vs. C2C12). Induction of noncoupled respiration revealed a significantly greater electron transfer capacity in C2C12 compared with L6 myoblasts, regardless of the substrate(s) provided. Mitochondrial metabolism was also investigated in differentiated L6 and C2C12 myotubes. Basal rates of oxygen consumption were not different between intact, adherent L6, and C2C12 myotubes; however, noncoupled respiration was significantly lower in L6 compared with C2C12 myotubes (P = 0.01). In summary, L6 myoblasts had lower respiration rates than C2C12 myoblasts, including lesser capacity for fatty acid oxidation and greater electron leak toward H2O2. L6 cells also retain a lower capacity for electron transfer compared with C2C12 following differentiation to form fused myotubes. Intrinsic differences in mitochondrial metabolism between these cell lines should be considered when modeling and investigating myocellular metabolism.
ISSN:0363-6143
1522-1563
DOI:10.1152/ajpcell.00343.2018