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X chromosome inactivation (XCI) is required for dosage compensation of X-linked genes in human female cells.Several researah groups have described the promiscuous XCI status in long-term cultured female human embryonic stem cells (hESCs), which impact their utility.Here, we traced the XCI dynamic changes during the hESCs derivation and early culture.The results showed the female initial hESCs (~Passage 5) expressed XCI marker XIST and H3K27me3 dot and had a random inactivated X chromosome.