Fig. 6

Comparison of PVA and FBS on hepatic differentiation. a Scheme for hepatic differentiation using 0.1% PVA or 1% FBS in the advanced protocol. OSM: Oncostatin M. b Comparison of morphological change during the differentiation when used 0.1% PVA or 10% FBS. Scale bar = 200 μm. c-e The mRNA levels of c hepatic progenitor markers (CPM and EpCAM), d liver-specific plasma and nuclear protein (AFP, ALB, HNF1A, and HNF4A), and e liver-specific metabolism (CYP1A2, CYP3A4, UGT1A6, MRP2, and ASGR1) in primary human hepatocytes (PHH), PVA, and FBS groups. GAPDH was used as an internal control. #, P < 0.05. f The expression of EpCAM, ALB, and CYP3A4 in day 0 (undifferentiated cells) and differentiated cells (hepatocyte-like cells; HLCs) on day 18 after the induction of differentiation. Green, ALB and CYP3A4; Red, EpCAM; Blue, DAPI. Scale bar = 100 μm. g Comparison of MHC class I expression between AM-MSCs and AM-HLCs with PVA. h Detection of human albumin secretion in PHH and AM-HLCs, which was derived from three independent AM-MSCs. Day 0 were indicated undifferentiated cells, and Day 18 were indicated differentiated HLCs. i Measurement of CYP3A4 activity using two experiments: Left, detection of 1-Hydroxymidazolam using LC-ESI/MS/MS systems; Right, detection of luciferin density using luminescence systems (PHH: Technical replicate, n = 3; AM-MSCs and AM-HLCs: Biological replicate, n = 3). n.d., non-detected j Time-dependent comparison of effects on CPM and EpCAM expression during hepatic differentiation. k Oxygen consumption rates (OCR) according to the day of hepatic differentiation. Olig: oligomycin, FCCP: carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone, AA: antimycin A, Rot: rotenone. l Changes of OCR values of basal and maximal respiration, spare respiratory capacity, and ATP production. OCR values were normalized by the DNA concentration. P-values < 0.05 were considered significant. *, P < 0.05; **, P < 0.01; ***, P < 0.001