In vitro hepatocytic differentiation of FL-MSCs. (a) Morphological changes noticed in differentiated FL-MSCs after 3-4 weeks of culture in hepatogenic differentiation medium as compared to undifferentiated cells. Magnification: 100x. (b) RT-qPCR gene expression analysis demonstrated the decrease in stemness markers and the upregulation of hepatocyte markers mostly at the last maturation step of the differentiation process which is in correlation with morphological changes. Results are expressed as fold change in differentiated versus undifferentiated FL-MSC. ALB (albumin encoding gene), MRP2 (multidrug resistance-associated protein-2 encoding gene), FOXA1 (Forkhead Box A1 encoding gene), ACTA2 (α-SM-actin encoding gene), CDH1 (cadherin-1 encoding gene), CXCR4 (chemokine (C-X-C motif) receptor 4 encoding gene), SOX9 (SRY-Related HMG-Box 9 encoding gene), and SOX17 (SRY-Related HMG-Box 17 encoding gene). (c) Immunoreactivity to human hepatic marker and transcription factor specific antibodies: α-fetoprotein (AFP), albumin, HNF (hepatocyte nuclear factor) 3β, HNF4α, HNF6, and HNF1α. Presented images are representative of at least 4 different experiments. Negative control: secondary antibody alone. Scale magnification: 400x (except for HNF1α (all cells analyzed); Alb and HNF3b in Huh-7: 200x).