Disease modeling following organoid-based expansion of airway epithelial cells

Evelien Eenjes, Sander van Riet, Andre A. Kroon, Annelies M. Slats, P. Padmini S.J. Khedoe, Anne Boerema-De Munck, Marjon Buscop-Van Kempen, Dennis K. Ninaber, Irwin K.M. Reiss, Hans Clevers, Robbert J. Rottier, Pieter S. Hiemstra*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

10 Citations (Scopus)


Air-liquid interface (ALI) cultures are frequently used in lung research but require substantial cell numbers that cannot readily be obtained from patients. We explored whether organoid expansion [three-dimensional (3D)] can be used to establish ALI cultures from clinical samples with low epithelial cell numbers. Airway epithelial cells were obtained from tracheal aspirates (TA) from preterm newborns and from bronchoalveolar lavage (BAL) or bronchial tissue (BT) from adults. TA and BAL cells were 3D-expanded, whereas cells from BT were expanded in 3D and 2D. Following expansion, cells were cultured at ALI to induce differentiation. The impact of cell origin and 2D or 3D expansion was assessed with respect to 1) cellular composition, 2) response to cigarette smoke exposure, and 3) effect of Notch inhibition or IL-13 stimulation on cellular differentiation. We established well-differentiated ALI cultures from all samples. Cellular compositions (basal, ciliated, and goblet cells) were comparable. All 3D-expanded cultures showed a similar stress response following cigarette smoke exposure but differed from the 2D-expanded cultures. Higher peak levels of antioxidant genes HMOX1 and NQO1 and a more rapid return to baseline, and a lower unfolded protein response was observed after cigarette smoke exposure in 3D-derived cultures compared to 2D-derived cultures. In addition, TA- and BAL-derived cultures were less sensitive to modulation by DAPT or IL-13 than BT-derived cultures. Organoid-based expansion of clinical samples with low cell numbers, such as TA from preterm newborns is a valid method and tool to establish ALI cultures.

Original languageEnglish
Pages (from-to)L775-L786
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Issue number4
Publication statusPublished - 11 Oct 2021

Bibliographical note

Funding Information:
This study was supported by a grant from The Netherlands Organization for Health Research and Development (ZonMw; Grant No. 114021508), SSWO project S14-12 (Sophia Foundation for Medical Research), and Lung Foundation Netherlands (

Publisher Copyright:
© 2021 the American Physiological Society.


Dive into the research topics of 'Disease modeling following organoid-based expansion of airway epithelial cells'. Together they form a unique fingerprint.

Cite this