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To examine the mechanism underlying the self-renewal of pluripotent stem cells, we subcutaneously transplanted an Oct4-GFP (Oct4 promoter driving a green fluorescent protein reporter) mouse embryonic stem cells (ESCs) into nude mice.The results demonstrated that some residuals of the ESCs maintained Oct4-expression and acquire malignancy during transplantation, from which an embryonic carcinoma cells (ECCs) line (EH-G5) was established.