Kyoto human embryonic stem cell 1 (Institute for Frontier Medical Sciences)

Grow cell colonies on an MEF feeder layer and culture in 1:1 Dulbecco’s modified Eagle’s medium (DMEM) and Ham’s nutrient mixture F-12 supplemented with 2-mercaptoethanol (0.1 mM), MEM non-essential amino acids, recombinant human basic FGF (5 ng/ml), and KSR (20%). Dissociate colonies in CTK dissociation solution consisting of trypsin (0.25%), collagenase IV (0.1%), KSR (20%), and CaCl2 (1mM) in PBS.