The nuclear receptor THRB facilitates differentiation of human PSCs into more mature hepatocytes

Cell Stem Cell. 2022 May 5;29(5):795-809.e11. doi: 10.1016/j.stem.2022.03.015. Epub 2022 Apr 21.


To understand the mechanisms regulating the in vitro maturation of hPSC-derived hepatocytes, we developed a 3D differentiation system and compared gene regulatory elements in human primary hepatocytes with those in hPSC-hepatocytes that were differentiated in 2D or 3D conditions by RNA-seq, ATAC-seq, and H3K27Ac ChIP-seq. Regulome comparisons showed a reduced enrichment of thyroid receptor THRB motifs in accessible chromatin and active enhancers without a reduced transcription of THRB. The addition of thyroid hormone T3 increased the binding of THRB to the CYP3A4 proximal enhancer, restored the super-enhancer status and gene expression of NFIC, and reduced the expression of AFP. The resultant hPSC-hepatocytes showed gene expression, epigenetic status, and super-enhancer landscape closer to primary hepatocytes and activated regulatory regions including non-coding SNPs associated with liver-related diseases. Transplanting the hPSC-hepatocytes resulted in the engraftment of human hepatocytes into the mouse liver without disrupting normal liver histology. This work implicates the environmental factor-nuclear receptor axis in regulating the maturation of hPSC-hepatocytes.

Keywords: 3D culture; epigenetics; hepatocytes differentiation and maturation; human pluripotent stem cells; nuclear receptors; pBAF; transcriptional regulation.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation
  • Chromatin* / metabolism
  • Hepatocytes* / metabolism
  • Humans
  • Mice
  • Polymorphism, Single Nucleotide
  • Receptors, Cytoplasmic and Nuclear / metabolism
  • Regulatory Sequences, Nucleic Acid


  • Chromatin
  • Receptors, Cytoplasmic and Nuclear