Figure 7.

Effect of inflammation on ADHLSC hepatocytic differentiation potential. (A) Primed ADHLSC acquired polygonal shape with granular cytoplasm after in vitro hepatocytic differentiation similar to the standard condition. Magnification: 200×. (B) RT-qPCR gene expression analysis demonstrated that inflammation did not alter the mRNA expression of mesenchymal markers except for slug and vimentin. Slug expression remains upregulated in differentiated 24 h-primed cells and both undifferentiated and differentiated 9-day primed cells while the expression of vimentin remains downregulated in non-differentiated 24 h and 9-day primed cells as well as differentiated 9-day primed cells. Furthermore, inflammation did not impact the upregulation of hepatocytic markers that normally occurs after hepatocytic differentiation (Albumin expression in differentiated 24 h primed cells and MRP2 expression in 9-day primed and differentiated cells) which is correlated with the morphological changes described above in Figure 7A. For treated and untreated differentiated cell groups, results are expressed as fold change in differentiated versus undifferentiated ADHLSC. *** denotes a p value < 0.001, ** p < 0.01, * p < 0.05 vs. non-primed and non-differentiated control, one-way ANOVA followed by Dunnett post-hoc test. CD54 (Intercellular Adhesion Molecule 1; ICAM-1), SOX9 (SRY-Related HMG-Box 9 encoding gene), Snail (SNAI1), Slug (SNAI2), VIM (Vimentin), ALB (Albumin), MRP2 (multi-drug resistance-associated protein-2 encoding gene). (C) Undifferentiated and differentiated ADHLSC from untreated and treated groups were incubated with IPA substrate and luciferase activity was measured. Results are expressed as the % of relative luminescence unit detected in the differentiated ADHLSC versus undifferentiated counterparts. Data shown are the mean ± SEM of three independent experiments (three different samples from different donors).