Multilineage communication regulates human liver bud development from pluripotency

Nature. 2017 Jun 22;546(7659):533-538. doi: 10.1038/nature22796. Epub 2017 Jun 14.


Conventional two-dimensional differentiation from pluripotency fails to recapitulate cell interactions occurring during organogenesis. Three-dimensional organoids generate complex organ-like tissues; however, it is unclear how heterotypic interactions affect lineage identity. Here we use single-cell RNA sequencing to reconstruct hepatocyte-like lineage progression from pluripotency in two-dimensional culture. We then derive three-dimensional liver bud organoids by reconstituting hepatic, stromal, and endothelial interactions, and deconstruct heterogeneity during liver bud development. We find that liver bud hepatoblasts diverge from the two-dimensional lineage, and express epithelial migration signatures characteristic of organ budding. We benchmark three-dimensional liver buds against fetal and adult human liver single-cell RNA sequencing data, and find a striking correspondence between the three-dimensional liver bud and fetal liver cells. We use a receptor-ligand pairing analysis and a high-throughput inhibitor assay to interrogate signalling in liver buds, and show that vascular endothelial growth factor (VEGF) crosstalk potentiates endothelial network formation and hepatoblast differentiation. Our molecular dissection reveals interlineage communication regulating organoid development, and illuminates previously inaccessible aspects of human liver development.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Aged
  • Cell Communication*
  • Cell Differentiation*
  • Cell Hypoxia
  • Cell Lineage*
  • Cell Movement
  • Endothelium / cytology
  • Epithelial Cells / cytology
  • Extracellular Matrix / metabolism
  • Female
  • Fetus / cytology
  • Hepatocytes / cytology
  • Humans
  • Liver / cytology*
  • Liver / embryology*
  • Male
  • Middle Aged
  • Organogenesis*
  • Organoids / cytology
  • Pluripotent Stem Cells / cytology
  • Sequence Analysis, RNA
  • Signal Transduction
  • Single-Cell Analysis
  • Tissue Culture Techniques / methods*
  • Vascular Endothelial Growth Factor A / metabolism
  • Young Adult


  • Vascular Endothelial Growth Factor A