Effect of Microenvironment on Differentiation of Human Umbilical Cord Mesenchymal Stem Cells into Hepatocytes In Vitro and In Vivo

Figure 3

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, , compared with 0 d, compared with 3 d, and 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.

(a)

(b)

(c)

(d)

(e)

(f)

(g)