2.3. Primary cultures of rat hepatocyte
Hepatocytes were isolated from male Wistar rat using methods of Berry and Friend (1969) as modified by Stacey and Priestly (1978) and Pramyothin (1986). Cell viability was estimated immediately after the isolation by Trypan blue exclusion test using the exclusion index of >90%. Hepatocytes were washed and suspended in Eagle's MEM supplemented with 10% inactivated fetal bovine serum, 10−8 M insulin, 10−6 M dexamethasone, penicillin (100 IU/l) and streptomycin (100 μg/ml). Cells were placed in 24-well culture plates (Greiner, Germany) with a cell density of 1 × 105 cells/0.8 ml at 37 °C, under carbogen (95% air, 5% CO2) atmosphere.