Comparative study on isolation and mitochondrial function of adult mouse and rat cardiomyocytes.
J Mol Cell Cardiol
; 136: 64-71, 2019 11.
Article
em En
| MEDLINE
| ID: mdl-31521710
ABSTRACT
BACKGROUND:
Cultured adult mouse and rat cardiomyocytes are the best and low-cost cell model for cardiac cellular physiology, pathology, drug toxicity screening, and intervention. The functions of mouse cardiomyocytes decline faster than rat cardiomyocytes in culture conditions. However, little is known about the difference of mitochondrial function between cultured mouse and rat myocytes. METHODS ANDRESULTS:
A large number of adult mouse and rat cardiomyocytes were comparative isolated using a simple perfusion system. Cardiomyocytes mitochondrial functions were measured after 2â¯h, 1â¯day, 2â¯days, 3â¯days, and 4â¯days culture by monitoring mitoflashes. We found that the mitochondrial function of mouse myocytes was remarkedly declined on the third day. Then, we focused on the third day cultured mouse and rat myocytes, comparatively analyzing the respiration function and superoxide generation stimulated by pyruvate/malate/ADP and the mitochondrial permeability transition pore (mPTP) opening induction. Mouse myocytes showed lower respiration and mitoflash activity, but without the change of maximum uncoupled respiration when compared with rat myocytes. Although the response to superoxide production stimulated by respiration substrates was slower than rat myocytes, the basal superoxide generation is faster than the rat. The faster mitochondrial reactive oxygen species (ROS) generation of mouse myocytes upon laser stimulation triggered the faster mPTP opening compared with the rat. Finally, antioxidant MitoTEMPO pretreatment preserved the mitochondrial function of mouse myocytes on the third day.CONCLUSIONS:
The mitochondrial function and stability are different between cultured mouse and rat cardiac myocytes beyond 3â¯days even though they both belong to Muridae. Mitochondrial ROS impairs the mitochondrial functions of mouse cardiomyocytes on the third day. Suppressing superoxide maintained the mitochondrial function of mouse myocytes on the third day.Palavras-chave
Texto completo:
1
Base de dados:
MEDLINE
Assunto principal:
Separação Celular
/
Miócitos Cardíacos
/
Mitocôndrias Cardíacas
Tipo de estudo:
Prognostic_studies
Limite:
Animals
Idioma:
En
Revista:
J Mol Cell Cardiol
Ano de publicação:
2019
Tipo de documento:
Article
País de afiliação:
China