The human constitutive androstane receptor promotes the differentiation and maturation of hepatic-like cells

Dev Biol. 2013 Dec 15;384(2):155-65. doi: 10.1016/j.ydbio.2013.10.012. Epub 2013 Oct 18.


Expression of the constitutive androstane receptor (CAR, NR1I3) is enriched in the mature mammalian liver and increasingly recognized for its prominent role in regulating a myriad of processes including biotransformation, chemical transport, energy metabolism and lipid homeostasis. Previously, we demonstrated that CAR levels were markedly enhanced during the differentiation of hepatic-like cells derived from hESCs, prompting the hypothesis that CAR contributes a key functional role in directing human hepatogenesis. Here we demonstrate that over-expression of CAR in human embryonic stem cells (ESCs), transduced by a lentiviral vector, accelerates the maturation of hepatic-like cells, with CAR over-expressing cells exhibiting a 2.5-fold increase in albumin secretion by day 20 in culture differentiation, and significantly enhanced levels of mRNA expression of several liver-selective markers, including hepatic transcription factors, plasma proteins, biotransformation enzymes, and metabolic enzymes. CAR over-expressing cells also exhibited enhanced CITCO-inducible CYP3A7 enzymatic activity. Knockdown of CAR via siRNA attenuated the differentiation-dependent expression programs. In contrast, expression levels of the pregnane X receptor (PXR), a nuclear receptor most similar to CAR in primary sequence, were negligible in human fetal liver tissues or in the differentiating hESCs, and stable over-expression of PXR in hepatic-induced hESCs failed to enhance expression of hepatic phenotype markers. Together, these results define a novel role for human CAR in hepatic lineage commitment.

Keywords: CAR; Drug metabolism; Hepatic differentiation; Lentivirus; PXR; hESCs; siRNA.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Base Sequence
  • Cell Differentiation / physiology*
  • Cell Line
  • Constitutive Androstane Receptor
  • Cytochrome P-450 Enzyme System / genetics
  • DNA Primers
  • Embryonic Stem Cells / cytology
  • Humans
  • Liver / cytology*
  • Polymerase Chain Reaction
  • RNA, Messenger / genetics
  • RNA, Small Interfering
  • Receptors, Cytoplasmic and Nuclear / physiology*
  • Stem Cells / cytology


  • Constitutive Androstane Receptor
  • DNA Primers
  • NR1I3 protein, human
  • RNA, Messenger
  • RNA, Small Interfering
  • Receptors, Cytoplasmic and Nuclear
  • Cytochrome P-450 Enzyme System