Effect of long-term cyclic mechanical load on protein synthesis and morphological changes in cultured Myocardial cells from neonatal rat

Abstract

We studied the effect of prolonged cyclic mechanical stimulation on protein synthesis and morphological changes in cultured cardiac myocytes isolated from 3-day-old neonatal rats to develop an in vitro model for cardiac hypertrophy induced by a mechanical load. Myocardial cells were cultured on deformative culture plates with M-199 culture medium in the presence or absence of fetal calf serum (FCS), and a cyclic mechanical load was applied for 2 hours up to 15 days. Mechanical stimulation for 2 hours increased the 14C-phenylalanine incorporation rate of myocardial cells, both in the absence and presence of FCS or when the myocardial cells were either beating or arrested with tetrodotoxin. The incorporation rate always increased by mechanical stimulation during 15 days of cell culture as compared to cells that sustained no mechanical stress. The cell size and protein content, which increased gradually with mechanical stimulation and reached a maximum at 10 days, were even greater in the presence of FCS. Within 10 days myocardial cells had aligned in the direction of the maximal cyclic mechanical load. In these cells electron microscopy revealed an increase in the number of myofilaments associated with the development of mitochondria as recognized in the adult myocardial cells. These results indicate that long-term cyclic mechanical loading of cultured myocardial cells may be a good in vitro model for the study of cardiac hypertrophy.