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Cells are positive for pluripotent markers and express alkaline phosphatase, as seen by staining. Karyotype: normal karyotype (46,XY) at earlier passages, but contain a subpopulation with trysomy 16 at late passages (47,XY+16).
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.