2.3. Isolation of primary rat hepatocytes

Pellet containing hepatocytes was rinsed with cold Hanks’ balanced salt solution (HBSS) and subsequently centrifuged (50× g, 5 min, 4°C). This washing procedure was performed three times. Hepatocytes above 75% viability (evaluated by trypan blue exclusion) were cultured in medium M1 (Table 2) plated in 0.1 mg/mL collagen‐coated petri dishes at a density of 109 000 cells/cm² and maintained at 37°C in a humidified atmosphere of 5% CO₂. After 4 hours, cells were rinsed twice with Dulbecco's phosphate‐buffered saline (DPBS) and the medium was replaced by medium M2 (Table 2).

Medium composition. Detailed supplements and concentrations for cell culture media