Fig 1: Immunocytochemistry staining showing the expression of characteristic stage-specific protein markers at major stages of HLC differentiation. (A) OCT4, SSEA4, TRA-1-60, and SOX2 for iPSCs. (B) SOX17 and FOXA2 for DE cells. HNF4A, AFP, ALB, CK18, A1AT, and CYP3A4 for HLCs derived from (C) Method 1 and (D) Method 2. Scale bar, 400 µm. (E) Quantification of fluorescence intensity of hepatic protein marker immunostaining. n=4, *P<0.05, **P<0.01, ***P<0.001 using a two tailed Student's t-test. HLC, hepatocyte-like cell; DE, definitive endoderm; iPSC, induced pluripotent stem cell.
Fig 2: Functional characterization of HLCs. (A) Secretion of serum proteins ALB, fibronectin and A1AT. (B) Urea synthesis and release upon incubation with different concentrations of NH4Cl. (C) Periodic acid-Schiff staining showing glycogen storage. Scale bar, 100 µm. (D) Basal activities of CYP1A, CYP2D6, CYP2B6, CYP3A, and CYP2C9. (E) Induction of CYP1A, CYP2D6, CYP2B6, CYP3A and CYP2C9, respectively, after treatment with OME or PB. For CYP activity, 1 unit (U)=1 pmol metabolite/mg protein/h, and the metabolites are acetaminophen, 1'-hydroxy bufuralol, hydroxy bupropion, 1'-hydroxy midazolam and 4'-hydroxy diclofenac for CYP1A, CYP2D6, CYP2B6, CYP3A, and CYP2C9, respectively. All values are presented as the mean ± standard deviation. n=4 for serum protein secretion and CYP activity, and n=3 for urea synthesis. *P<0.05, **P<0.01, ***P<0.001. HLC, hepatocyte-like cell; ALB, albumin; A1AT, a-1-antitrypsin; CYP, cytochrome P450; OME, omeprazole; PB, phenobarbital; HLC_M1, HLCs derived from method 1; HLC_M2, HLCs derived from method 2.
Supplier Page from Abcam for Human alpha 1 Antitrypsin ELISA Kit, Fluorescent