Composition	Volume	Final concentration
DMEM/F-12	195 ml	78%
KO serum replacement	50 ml	20%
100× NEAA solution (10 mM)	2.5 ml	100 µM
L-glutamine/2-mercaptoethanol solution (100 mM)	2.5 ml	1 mM
Basic FGF (100 µg/ml) for hESCs culture	10 µl	4 ng/ml

Sterilize the medium by filtration using a 0.22 µm membrane filter and store for up to 2 weeks at 2–8°C.