Evaluation of chloroform cardiotoxicity utilizing a modified isolated rat cardiac myocytes

Abstract

An improved technique for the isolation of cardiac myocytes from adult rat is described. This method produced a high yield of isolated cardiomyocytes (30–35 × 10 6 cells/heart) with viability over 88%. The rod-shaped cells represented about 70% of the viable cardiac myocytes. The leakage of creatine phosphokinase (CPK) in the incubation medium from the control was about 37% at zero time. This model was used to investigate chloroform (CHCl 3) cardiotoxicity by measuring dye exclusion, change the viable cardiac myocytes shape and CPK leakage. As a result of CHCl 3 exposure, there was a significant decrease in Trypan blue exclusion after 30 min and during the entire time course. The percentage of rod-shaped cells decreased significantly during the whole period of the study. Also, a significant elevation in CPK. leakage after treatment with 1000 ppm CHCl 3 was observed. The data show that the present technique for cardiomyocytes isolation from adult rats produced high percentage of viable cells. Furthermore, the toxic effect of CHCl 3 on cardiac myocytes was time dependent.