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Provided is a chemically defined medium for eukaryotic cell culture, comprising water, at least one carbon source, one or more vitamins, one or more salts, one or more fatty acids, one or more buffer components, selenium, at least one substance of the group of Functional Inhibitors of Acid Sphingomyelinase (FIASMAs) and at least one polypeptide of TGF-β superfamily. The medium according to this invention enables continuous proliferation and serial passageability of human pluripotent stem cells.