Figure 1.
Differentiation of Hepatocyte-like Cells from Human Pluripotent Stem Cells
(A) Schematic description of the four stages of hepatocyte differentiation.
(B) Undifferentiated hESCs. hESCs were positive for NANOG, SOX2, OCT4, and SSEA4 and maintained in the pluripotent state.
(C) Induction of definitive endoderm. The expression of specific marker genes SOX17 and CXCR4 were induced in hESC-derived definitive endoderm at day 3.
(D) Hepatic specification. At day 6 after differentiation, the differentiated hepatic progenitors were AFP and CK19 double-positive and homogenously expressed the hepatic gene HNF4A.
(E) Maturation of hepatic progenitors into hepatocyte-like cells. The HLCs underwent functional maturation from day 7. At day 17, the resulting cells expressed specific genes unique to hepatocytes, including AFP, ALB, A1AT, NTCP, and coagulation factor F5. Lipid droplets were formed in these cells.
(F) Induction of genes preferentially expressed in the liver during differentiation. Heatmap of RNA-seq expression values (log10(1 + FPKM)) of the 291 RefSeq genes preferentially expressed (Liu et al., 2008) in liver at day 5, day 10, and day 12 of differentiation.