Evaluation of chloroform cardiotoxicity utilizing a modified isolated rat cardiac myocytes

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 x 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 (CHCl3) cardiotoxicity by measuring dye exclusion, change the viable cardiac myocytes shape and CPK leakage. As a result of CHCl3 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 CHCl3 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 CHCl3 on cardiac myocytes was time dependent.