In vitro generation of human pluripotent stem cell derived lung organoids

Elife. 2015 Mar 24:4:e05098. doi: 10.7554/eLife.05098.

Abstract

Recent breakthroughs in 3-dimensional (3D) organoid cultures for many organ systems have led to new physiologically complex in vitro models to study human development and disease. Here, we report the step-wise differentiation of human pluripotent stem cells (hPSCs) (embryonic and induced) into lung organoids. By manipulating developmental signaling pathways hPSCs generate ventral-anterior foregut spheroids, which are then expanded into human lung organoids (HLOs). HLOs consist of epithelial and mesenchymal compartments of the lung, organized with structural features similar to the native lung. HLOs possess upper airway-like epithelium with basal cells and immature ciliated cells surrounded by smooth muscle and myofibroblasts as well as an alveolar-like domain with appropriate cell types. Using RNA-sequencing, we show that HLOs are remarkably similar to human fetal lung based on global transcriptional profiles, suggesting that HLOs are an excellent model to study human lung development, maturation and disease.

Keywords: developmental biology; endoderm; foregut; human; lung; organoids; pluripotent stem cells; spheroid; stem cells.

Publication types

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

MeSH terms

  • Cell Culture Techniques / methods
  • Cell Differentiation / genetics
  • Cell Line
  • Cells, Cultured
  • Embryonic Stem Cells / cytology
  • Embryonic Stem Cells / metabolism
  • Endoderm / cytology
  • Endoderm / metabolism
  • Gene Expression Profiling
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism
  • Lung / cytology*
  • Lung / embryology
  • Lung / metabolism
  • Microscopy, Confocal
  • Microscopy, Electron, Transmission
  • Organogenesis*
  • Organoids / cytology*
  • Organoids / metabolism
  • Organoids / ultrastructure
  • Pluripotent Stem Cells / cytology*
  • Pluripotent Stem Cells / metabolism
  • Reproducibility of Results
  • Reverse Transcriptase Polymerase Chain Reaction
  • Spheroids, Cellular / cytology
  • Spheroids, Cellular / metabolism
  • Tissue Engineering / methods