Mitochondrial remodelling is really a central feature of stem cell differentiation. However, little is famous concerning the regulatory mechanisms over these processes. Formerly, we discovered that a medicinal inhibitor of glycogen synthase kinase-3|á and -3|, CHIR-99021, initiates human adipose stem cell differentiation into human definitive endodermal progenitor cells (hEPCs), that have been forwarded to differentiate synchronously into hepatocyte-like cells after further treatment with mixtures of soluble factors. Within this study, we reveal that CHIR-99021 promotes mitochondrial biogenesis, the expression of PGC-1|á (also referred to as PPARGC1A), TFAM and NRF1 (also referred to as NFE2L1), oxidative phosphorylation capacities, and producing reactive oxygen species in hEPCs. Blocking mitochondrial dynamics using siRNA targeting DRP1 (also referred to as DNM1L) impaired definitive endodermal differentiation. Downregulation of |-catenin (CTNNB1) expression weakened the result of CHIR-99021 around the induction of mitochondrial remodelling and also the expression of transcription factors for mitochondrial biogenesis. Furthermore, CHIR-99021 decreased the expression of miR-19b-2-5p, miR-23a-3p, miR-23c, miR-130a-3p and miR-130a-5p in hEPCs, which target transcription factors for mitochondrial biogenesis. These data show CHIR-99021 plays a part in mitochondrial structure and performance remodelling via activation from the |?-catenin signalling path and inhibits the expression of miRNAs during definitive endodermal differentiation.This information has an connected First Person interview using the first author from the paper.