Production of 3-D Airway Organoids From Primary Human Airway Basal Cells and Their Use in High-Throughput Screening

Curr Protoc Stem Cell Biol. 2016 May 12;37:IE.9.1-IE.9.15. doi: 10.1002/cpsc.1.

Abstract

The ability of human airway basal cells to serve as progenitor cells in the conducting airway makes them an attractive target in a number of respiratory diseases associated with epithelial remodeling. This unit describes a protocol for the culture of 'bronchospheres', three-dimensional (3-D) organoids that are derived from primary human airway basal cells. Mature bronchospheres are composed of functional multi-ciliated cells, mucin-producing goblet cells, and airway basal cells. In contrast to existing methods used for the culture of well-differentiated human airway epithelial cells, bronchospheres do not require growth on a permeable support and can be cultured in 384-well assay plates. The system provides a mechanism for investigating the regulation of basal cell fate during airway epithelial morphogenesis, as well as a basis for studying the function of the human airway epithelium in high-throughput assays. © 2016 by John Wiley & Sons, Inc.

Keywords: bronchosphere; ciliated cell; goblet cell; high throughput screening; mucociliary.

MeSH terms

  • Bronchi / cytology*
  • Cell Proliferation
  • Cells, Cultured
  • High-Throughput Screening Assays / methods*
  • Humans
  • Imaging, Three-Dimensional*
  • Nucleic Acid Hybridization
  • Organoids / cytology*
  • RNA / isolation & purification
  • Reproducibility of Results
  • Spheroids, Cellular / cytology
  • Tissue Culture Techniques / methods*

Substances

  • RNA