Fig 1: Detection of secreted albumin and blood urea nitrogen by ELISA. The ALB and BUN concentration of UC-MSCs and BM-MSCs culture supernatants collected on weeks 1, 2, 3, 4 and 5 following hepatic differentiation were measured. The medium of hepatocyte was measured as a control. Each bar represents the mean ± standard deviation (n=3). *P<0.05 vs. BM-MSCs. UC-MSCs, umbilical cord mesenchymal stem cells; BM-MSCs, bone marrow derived mesenchymal stem cells; W, weeks; ALB, albumin; BUN, blood urea nitrogen.
Fig 2: Immunofluorescent analysis of hepatocyte-specific proteins. (A) UC-MSCs and (B) BM-MSCs were examined for their expression of (A1 and B1) ALB, (A3 and B3) AFP, and (A5 and B5) CYP3A4 following hepatic differentiation for 4 weeks. (A2, 4 and 6, and B2, 4 and 6) Cells cultured in the growth medium were as negative controls. Scale bars, 100 µm. UC-MSCs, umbilical cord mesenchymal stem cells; BM-MSCs, bone marrow derived mesenchymal stem cells; ALB, albumin; AFP, a-fetoprotein; CYP3A4, cytochrome P450 3A4.
Fig 3: Model for the regulation of ML141-induced hepatocyte differentiation. In hADSCs, ML141 induced the hepatocyte differentiation by a mechanism involving the Wnt5a/PI3K/miR-122 signaling pathway, and regulated positively the hepatic specific genes and function, importantly the exosome release. ALB albumin, HNF hepatocyte nuclear factor, NSC NSC5476, PI3K phosphatidylinositol-3 kinase, WRT Wortmannin
Fig 4: Protein levels of ALB, TAT, CYP3A4, G-6P, a1AT and AFP were analyzed by western blot analysis. *P<0.05 vs. BM-MSCs. UC-MSCs, umbilical cord mesenchymal stem cells; BM-MSCs, bone marrow derived mesenchymal stem cells; ALB, albumin; CYP3A4, cytochrome P450 3A4; TAT, tyrosine-aminotransferase; G-6P, glucose-6phosphate; a1AT, a1 antitrypsin; AFP, a-fetoprotein.
Fig 5: Effect of CUDR overexpression on hepatic differentiation. (A) RT-qPCR analysis of CUDR expression levels in HuMSCs transduced with lentiviruses containing either CUDR or negative control. (B) ALB and BuN concentration of the supernatant were analyzed by ELISA. (C) Hepatic differentiation was evaluated with reverse transcription-quantitative PCR analysis of ALB, AFP, CYP3A4, G6P, TAT and a1AT. (D) Comparison of ALB, CYP3A4 and AFP protein between HuMSCs transfected with CUDR and cells cultured in medium containing cytokines via immunocytochemistry. *P<0.05 vs. control group. All experiments were performed three independent times. HuMSCs, human umbilical cord mesenchymal stem cells; ALB, albumin; AFB, a fetoprotein; CYP3A4, cytochrome P450 3A4; CUDR, cancer up-regulated drug resistant; BuN, blood urea nitrogen; G6P, glucose-6-phosphatase; TAT, tyrosine aminotransferase; a1AT, a1-antitrypsin.
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