TSC media

Reagent	Final concentration	Amount
RPMI1640	1×	385
FBS – ES Cell Qualified (not heat inactivated)	20% (v/v)	100 mL
Penicillin-Streptomycin	1% (v/v)	5 mL
2-mercaptoethanol	100 μM	900 μL
Glutamax	2 mM	5 mL
Na-Pyruvate	1 mM	5 mL
Total		500 mL
TSC basal media (above)	1×	50 mL
FGF4	25 ng/mL	12.5 μL
Heparin	1 μg/mL	25 μL
Total	n/a	50 mL

Filter sterile with a 0.22 μm filter and store without growth factors up to 4 weeks at 4°C. Store with growth factors for up to one week at 4°C