Fig. 3
Characterization of NTCP mRNA expression in D-UCMSCs. a Sodium-taurocholate cotransporting peptide (NTCP) mRNA expression: hepatogenic differentiation induces NTCP mRNA modulation in differentiated umbilical cord mesenchymal stem cells (D-UCMSCs; 60.0 ± 44.5 SD, n = 5) as compared to UCMSCs (1.6 ± 1.2 SD, n = 3; p = 0.0357 by Mann-Whitney U test, two-tailed). The relative quantification method was normalized using TBP and POP4 reference genes. b NTCP expression at Step 3 of differentiation: the third step of hepatogenic differentiation induces NTCP mRNA expression (5.9 ± 0.2 SD) as compared to Step 2 (0.6 ± 0.2 SD) and Step 1 (0.2 ± 0.7 SD). Values are expressed as fold increase compared to UCMSCs (p = 0.0047 by unpaired t test, two-tailed; n = 3). c NTCP mRNA expression post-hepatogenic differentiation: maintenance medium containing dexamethasone (Dexa) 10–6 M and HGF 10 ng/ml was added at the end of the hepatogenic differentiation protocol. The NTCP mRNA expression kinetic was sustained and increased at day 4 post-differentiation (p = 0.05 by Mann-Whitney U test, one-tailed; n = 3). Values are expressed as ΔΔ Ct fold increase as compared to day 0 (end of differentiation). d The effect of Dexa on NTCP mRNA expression: D-UCMSCs were treated with Dexa (10 nM, 100 nM, and 1000 nM) at Step 3 of hepatogenic differentiation. NTCP mRNA levels were measured by RT-qPCR (n = 2). NTCP expression was increased in a dose-dependent manner by Dexa (6.5 ± 1.6 SD 10 nM; 8.4 ± 0.6 SD 100 nM; and 16.1 ± 1.2 SD 1000 nM). Values are expressed as ΔΔ Ct fold increase versus Step 2 (end of differentiation). e Inhibition of NTCP mRNA expression in D-UCMSCs treated by RU486. 1 μM RU486 was added at the end of hepatogenic differentiation for 4 days with Dexa 10–6 M. qPCR analysis revealed that NTCP mRNA expression was significantly reduced by treatment with 1 μM RU486 as well as CYP3A4 mRNA expression (p < 0.001 by multiple t test). Values expressed as relative quantity compared to UCMSCs