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This chapter introduces a new method of maintaining human embryonic stem cells (hESCs) in the undifferentiated state through treatment with a GSK-3 inhibitor, BIO, under a feeder-free condition. Additionally, methods are introduced that determine multidifferentiation potential of hESCs by differentiating into a specific type or random heterogenous cell populations in vitro. These approaches will become a fundamental platform to identify molecular networks regulating cell fate determination in hESCs and mouse embryonic stem cells.
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