Figure 5
From: Direct conversion of porcine primary fibroblasts into hepatocyte-like cells
Morphology, Albumin secretion and gene expression analysis of 3TFs-piHeps. (A) Bright field pictures of PKF cells undergoing directed hepatic conversion at days 7 and 14 post-transduction (p.t.) with 3TFs CEBPα, FOXA1 and HNF4α2, 12TFs or Neg-Ctrl. Blue scale bar equals to 100 µm. (B) Albumin secretion of 3TF-piHeps and 12TF-piHeps supernatants compared to PPHs supernatant diluted 1:10. (C) Expression of fibroblastic, endodermal, hepatoblastic and mature hepatic markers in PKF transduced with 3TFs: CEBPα, FOXA1 and HNF4α2, 12TFs or Neg-Ctrl, as well as PPHs and Fetal liver samples. All data from qPCR of 3TF vs 12TFs vs Neg-Ctrl as well as Albumin ELISA were analyzed using one-way ANOVA, with Tukey’s post-test. qPCR data of PPHs vs Fetal liver samples were analyzed with multiple comparisons t-test. Significance from n = 3 independent values is displayed as *p < 0.05; ****p ≤ 0.0001. Genes GATA4, GATA6, HNF1α, HNF1β were analyzed only in 3TFs-piHeps and Neg-Ctrl, but not in 12TFs-piHeps, since the CDS of each lentiviral vector present in the 12TF pool would also be detected.
