The present invention relates to self-renewing liver organoids which express a specific liver-specific gene. Liver organoids according to the present invention exhibit characteristics of liver cells that are more mature than 2D-differentiated liver cells, can be subcultured up to at 67 times or more, and exhibit self-renewability, wherein the characteristics of mature liver cells are maintained even after repeated subculturing. Thus, the liver organoids can be effectively used for: predicting toxicity, regenerative response, and inflammatory response; drug screening; and modeling diseases such as liver steatosis.
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