Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2016 May 12:37:IE.9.1-IE.9.15.
doi: 10.1002/cpsc.1.

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

Marc Hild  1 , Aron B Jaffe  1
Affiliations

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

Marc Hild et al. Curr Protoc Stem Cell Biol. .

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.

PubMed Disclaimer

References

Literature Cited

    1. Araya, J. , Cambier, S. , Markovics, J.A. , Wolters, P. , Jablons, D. , Hill, A. , Finkbeiner, W. , Jones, K. , Broaddus, V.C. , Sheppard, D. , Barzcak, A. , Xiao, Y. , Erle, D.J. , and Nishimura, S.L. 2007. Squamous metaplasia amplifies pathologic epithelial-mesenchymal interactions in COPD patients. J. Clin. Invest. 117:3551-3562. doi: 10.1172/JCI32526.
    1. Danahay, H. , Pessotti, A.D. , Coote, J. , Montgomery, B.E. , Xia, D. , Wilson, A. , Yang, H. , Wang, Z. , Bevan, L. , Thomas, C. , Petit, S. , London, A. , LeMotte, P. , Doelemeyer, A. , Velez-Reyes, G.L. , Bernasconi, P. , Fryer, C.J. , Edwards, M. , Capodieci, P. , Chen, A. , Hild, M. , and Jaffe, A.B. 2015. Notch2 is required for inflammatory cytokine-driven goblet cell metaplasia in the lung. Cell Rep. 10:239-252. doi: 10.1016/j.celrep.2014.12.017.
    1. Debnath, J. , Muthuswamy, S.K. , and Brugge, J.S. 2003. Morphogenesis and oncogenesis of MCF-10A mammary epithelial acini grown in three-dimensional basement membrane cultures. Methods 30:256-268. doi: 10.1016/S1046-2023(03)00032-X.
    1. Fulcher, M.L. , Gabriel, S. , Burns, K.A. , Yankaskas, J.R. , and Randell, S.H. 2005. Well-differentiated human airway epithelial cell cultures. Methods Mol. Med. 107:183-206. doi: 10.1385/1-59259-861-7:183.
    1. Gray, T.E. , Guzman, K. , Davis, C.W. , Abdullah, L.H. , and Nettesheim, P. 1996. Mucociliary differentiation of serially passaged normal human tracheobronchial epithelial cells. Am. J. Respir. Cell Mol. Biol. 14:104-112. doi: 10.1165/ajrcmb.14.1.8534481.

LinkOut - more resources