F-12 Medium Supplemented with
Reagent	Stock Solution	Final Solution	Amount
Fetal Calf serum		10%	50 ml
Sodium pyruvate	100 mM	1 mM	5 ml
L-Glutamine	200 mM	2 mM	5 ml
Hepes	1 M	10 mM	5 ml
Gentamicin	50 mg/ml	50 μg/ml	500 μl
Penicillin/streptomycin	10,000 U/ml/10 mg/ml	100 U/ml/100 μg/ml	5 ml
Insulin	5 mg/ml	5 μg/ml	500 μl
T3	2 x 10- 8 M	2 x 10-11 M	500 μl
Adenine	1.8 x 10- 1 M	1.8 x 10- 3 M	5 ml
Transferrin	5 mg/ml	5 μg/ml	500 μl
heparin	50 mg/ml	0.1 mg/ml	1 ml
ECGS	3 mg/ml	3 μg/ml	5 ml
bFGF	25 μg/ml	5 ng/ml	100 μl
SCF	10 μg/ml	5 ng/ml	250 μl
HGF	5 μg/ml	2 ng/ml	200 μl
Endothelin 3	50 μg/ml	10 ng/ml	200 μl
LIF	20 μg/ml	4 ng/ml	100 μl
VEGF	5 μg/ml	1 ng/ml	100 μl
Choleran toxin	10-7 M	10-10 M	500 μl
Note: The amount cited above is for preparation of 500 ml of 3-D culture media.  Media must be stored at 4 °C for no more than 2 weeks