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. Author manuscript; available in PMC: 2017 May 1.
Published in final edited form as: Stem Cell Res. 2016 Mar 29;16(3):640–650. doi: 10.1016/j.scr.2016.03.009

Figure 4. Hepatic specification and maturation of DE cells passed and differentiated in wells of a 96-well plate.

Figure 4

(A) After 8 days of hepatic specification (day 12), the cells were confluent, but did not display a hepatocyte morphology (phase contrast, magnification: 10x). Cells were positive for HNF4A and AFP, but remained negative for albumin, as assessed by immunofluorescence (Magnification: 20x).

(B) After hepatic maturation (day 15), hepatic-like cells (HLCs) displayed typical polygonal morphology (phase contrast, magnification: 10x) and were positive for AFP, albumin and HNF4A (magnification: 20x).

(C) Activity of the cytochrome P450 3A family (Substrate: Luc-PFBE) and CYP3A4 isoform (Substrate: Luc-IPA) in HLCs compared to hPSCs.

(D) Hep-Par1 immunohistochemistry visualization of engrafted HLCs 14 days after intrasplenic injection of 4 million HLCs in the spleen of MUP-uPA/SCID/Bg mice.

See Supplemental Figure 3 for more functional characterization of HLCs and assessment of intra-hepatic engraftment.