Table 1.

Medium composition for primary rat hepatocyte plating and maintenance.

Component	Volume	Final concentration	Cat. No. (manufacturer)
a. Hepatocyte plating medium composition
Williams' E Medium	500 mL	–	#A1217601 (Gibco, USA)
Fetal Bovine Serum	25 mL	5%	#16000044 (Gibco, USA)
GlutaMAX (100X)	5 mL	2 mM	#35050061 (Gibco, USA)
HEPES pH 7.4 (1 M)	7.5 mL	15 mM	H3375 (Sigma, USA)
Penicillin-Streptomycin	5 mL	1X	#10378016 (Gibco, USA)
ITS-G (100X)	5 mL	1X	#41400045 (Gibco, USA)
Nicotinamide (5 M)	0.2 mL	0.2 mM	N0636 (Sigma, USA)
Dexamethasone (10 mM)	0.5 mL	1 μM	#041-18861 (Wako, Japan)
mEGF	0.25 mL	20 ng/mL	SRP3196 (Sigma, USA)
Ascorbic Acid (0.5 M)	0.5 mL	0.5 mM	#013-12061 (Wako, Japan)
b. Hepatocyte maintenance medium composition
Williams' E Medium	500 mL	–	#A1217601 (Gibco, USA)
Sodium Pyruvate (100X)	5 mL	1 mM	#11360070 (Gibco, USA)
GlutaMAX (100X)	5 mL	2 mM	#35050061 (Gibco, USA)
HEPES pH 7.4 (1 M)	7.5 mL	15 mM	H3375 (Sigma, USA)
Penicillin-Streptomycin	2.5 mL	0.5X	#10378016 (Gibco, USA)
ITS+ (100X)	5 mL	1X	#354352 (Corning, USA)
Nicotinamide (5 M)	0.2 mL	0.2 mM	N0636 (Sigma, USA)
Dexamethasone (10 mM)	0.05 mL	0.1 μM	#041-18861 (Wako, Japan)
mEGF	0.25 mL	20 ng/mL	SRP3196 (Sigma, USA)
Ascorbic Acid (0.5 M)	0.5 mL	0.5 mM	#013-12061 (Wako, Japan)