Primary Human Cardiomyocyte Culture: A model for assessment of cardiovascular disorders

Abstract

The purpose of this research is to establish an in vitro cardiac system using primary human cardiomyocyte cultures (PHCC). We hypothesized that norepinephrine (NE) and endothelin‐1 (ET‐1) will contribute to human cardiomyocyte growth. PHCC were obtained from Celprogen, San Pedro, CA and grown with varied concentrations of NE or ET‐1 for 72h. Protein expressions of contractile proteins were analyzed via Immunoblot and immunocytochemistry. Analysis of confocal images showed a significant decrease in nuclear area in all NE treatment groups. NE (10 μmol) stimulated significant increases in fluorescent intensities of the contractile proteins, f‐actin, troponin and tropomyosin. The protein expression of myosin light chain was significantly lower with increasing concentrations of NE. In addition, the average cell area increased in all groups of ET‐1. ET‐1 (1, 10, 100 nmol) exhibited a significant decrease of fluorescent intensities of f‐actin, troponin and tropomyosin. PHCC were also grown in the presence of LPS (1, 10, 100μg/ml) in a 6‐well plate for 72h. Viability data showed the cells recovered from the endotoxin treatment and maintain steady growth in each concentration of LPS. In contrast to NE, ET‐1 exhibited a decrease on the expression of contractile proteins in PHCC.