Journal: BioMed Research International
Article Title: Effect of Microenvironment on Differentiation of Human Umbilical Cord Mesenchymal Stem Cells into Hepatocytes In Vitro and In Vivo
doi: 10.1155/2016/8916534
Figure Lengend Snippet: Determination of hepatocyte markers and function after differentiation in vitro . ((a) and (b)) Determination of hepatocyte markers by western blotting. (c) Typical high-performance liquid chromatography/mass spectrometry chromatograms of midazolam and diazepam (internal standard) in culture media samples. (1) Blank culture medium + diazepam; (2) blank culture medium + midazolam (500 ng/mL) + diazepam; (3) sample culture medium after 2 h incubation with hUCMSCs induced by LHS. (d) Linear curve for midazolam. (e) Effects of LHS on CYP3A enzyme activity in hUCMSCs after 3, 5, and 7 days. (f) Effects of LHS on blood urea nitrogen production in hUCMSCs after 3, 5, and 7 days. (g) Effects of LHS on ALB externalization in hUCMSCs after 3, 5, and 7 days (mean ± SD, n = 6, ∗ P < 0.01 compared with 0 d, # P < 0.05 compared with 3 d, and △ P < 0.05 compared with 5 d). hUCMSC, human umbilical cord-derived mesenchymal stem cell; NC, negative control group; PC, positive control group of QSG-7701 human hepatic cell line; 0 d, undifferentiated hUCMSCs; 3 d, 5 d, 7 d, 3 d, 5 d, and 7 days after induction by LSH, respectively; AFP, α -fetoprotein; CK18, cytokeratin 18; TPH2, tryptophan 2,3-dioxygenase; LHS, liver homogenate supernatant.
Article Snippet: Liver slides were then incubated with diluted primary antibodies against anti-human CK18 (1 : 100, Abcam, Cambridge Science Park, Cambridge, UK), human AFP (1 : 50), human TPH2 (1 : 20), and human ALB (1 : 50) (Cell Signaling Technology, Inc., Danvers, MA, USA), followed by biotin-conjugated secondary antibody.
Techniques: In Vitro, Western Blot, High Performance Liquid Chromatography, Mass Spectrometry, Incubation, Activity Assay, Derivative Assay, Negative Control, Positive Control