RT Journal Article
SR Electronic
T1 Disease modelling following organoid-based expansion of airway epithelial cells
JF European Respiratory Journal
JO Eur Respir J
FD European Respiratory Society
SP 4322
DO 10.1183/13993003.congress-2020.4322
VO 56
IS suppl 64
A1 Pieter S. Hiemstra
A1 Evelien Eenjes
A1 Sander Van Riet
A1 Andre A. Kroon
A1 Annelies M. Slats
A1 Irwin K.M. Reiss
A1 Hans Clevers
A1 Robbert J. Rottier
YR 2020
UL http://erj.ersjournals.com/content/56/suppl_64/4322.abstract
AB Air-liquid interface (ALI) epithelial cell (EC) cultures are useful for studying the effect of exposure to airborne substances, but require substantial cell numbers that cannot always obtained from patients. Recent studies show that 3D organoid cultures can be initiated from samples with limited EC numbers, such as BAL (Sachs et al, 2019). Aim of the present study was to compare expansion of EC using 3D organoid culture to conventional 2D plastic-basic expansion for establishment of ALI cultures, and whether such cultures can be used for exposure to cigarette smoke (CS). Airway EC were obtained from tracheal aspirates (TA) from preterm newborns and bronchoalveolar lavage (BAL), or from bronchial tissue (BT) from adults. TA and BAL cells were 3D-expanded, whereas cells from BT were expanded in 3D and 2D. Following expansion, EC were cultured at ALI to induce differentiation. The impact of cell origin and 2D or 3D expansion was assessed with respect to (i) cellular composition; (ii) response to CS exposure; (iii) effect of Notch inhibition or IL-13 stimulation on cellular differentiation. Well-differentiated ALI cultures were established from all samples and cellular compositions were comparable. All 3D-expanded cultures showed a similar oxidative stress and unfolded protein response following CS exposure, but different from the 2D-expanded cultures. Additionally, TA- and BAL-derived cultures were less sensitive to modulation by DAPT or IL-13 than BT-derived cultures. We conclude that organoid-based expansion of clinical samples with low epithelial cell numbers, such as tracheal aspirates from preterm newborns, is a valid method and tool to establish ALI cultures.FootnotesCite this article as: European Respiratory Journal 2020; 56: Suppl. 64, 4322.This abstract was presented at the 2020 ERS International Congress, in session “Respiratory viruses in the "pre COVID-19" era”.This is an ERS International Congress abstract. No full-text version is available. Further material to accompany this abstract may be available at www.ers-education.org (ERS member access only).