Fig. 5

An efficient protocol for enhancing the hepatic differentiation potential of AM-MSCs. a The conventional and advanced in vitro hepatic differentiation protocols. The advanced protocol was modified from the conventional protocol by adding several chemicals highlighted in blue. OSM: Oncostatin M. b Morphology of cells during hepatic differentiation and ALB expression on day 14. Green, ALB; Blue, DAPI. Scale bar = 200 μm. c Fluorescence intensity of detected albumin in (b). d Hierarchical cluster analysis, and heatmap of endoderm-related transcriptomes in AM-MSCs (n = 3) and UCM-MSCs (n = 3). e Numbers of endoderm-related genes differentially expressed between AM-MSCs and UCM-MSCs. f Heat map of hepatic development-associated gene expression in AM-MSCs (n = 3) and UCM-MSCs (n = 3). g RT-qPCR analysis of GATA6 in AM-MSCs and UCM-MSCs. h Scheme of the effect of GSK3 inhibitor (CHIR99021) on AM-MSCs. i Regulation of GATA6, and SOX17 in AM-MSCs in the presence of CHIR99021. j Effect of the different protocol on the regulation of GATA6 and SOX17 in AM-MSCs. k RT-qPCR analysis of selected hepatic progenitor markers. l Expression of early hepatic (AFP) and hepatic maturation (ALB, and HNF4A) markers. GAPDH was used as an internal control for RT-qPCR. Two-way analysis of variance (ANOVA) was performed on the data for differentiation days 0, 7, and 14 points. P-values < 0.05 were considered significant. *** P < 0.001. Conv. Conventional protocol; Adv. Advanced protocol