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Mastalerz, M. ; Dick, E. ; Chakraborty, A. ; Hennen, E. ; Schamberger, A.C. ; Schröppel, A. ; Lindner, M.* ; Hatz, R.A.* ; Behr, J.* ; Hilgendorff, A. ; Schmid, O. ; Staab-Weijnitz, C.A.

Validation of in vitro models for smoke exposure of primary human bronchial epithelial cells.

Am. J. Physiol. Lung Cell Mol. Physiol. 322, L129-L148 (2021)
Postprint Research data DOI PMC
RATIONALE: The bronchial epithelium is constantly challenged by inhalative insults including cigarette smoke (CS), a key risk factor for lung disease. In vitro exposure of bronchial epithelial cells using CS extract (CSE) is a widespread alternative to whole CS (wCS) exposure. However, CSE exposure protocols vary considerably between studies, precluding direct comparison of applied doses. Moreover, they are rarely validated in terms of physiological response in vivo and the relevance of the findings is often unclear. METHODS: We tested six different exposure settings in primary human bronchial epithelial cells (phBECs), including five CSE protocols in comparison with wCS exposure. We quantified cell-delivered dose and directly compared all exposures using expression analysis of 10 well-established smoke-induced genes in bronchial epithelial cells. CSE exposure of phBECs was varied in terms of differentiation state, exposure route, duration of exposure, and dose. Gene expression was assessed by quantitative Real-Time PCR (qPCR) and Western Blot analysis. Cell type-specific expression of smoke-induced genes was analyzed by immunofluorescent analysis. RESULTS: Three surprisingly dissimilar exposure types, namely chronic CSE treatment of differentiating phBECs, acute CSE treatment of submerged basal phBECs, and wCS exposure of differentiated phBECs performed best, resulting in significant upregulation of seven (chronic CSE) and six (acute wCS, acute submerged CSE exposure) out of 10 genes. Acute apical or basolateral exposure of differentiated phBECs with CSE was much less effective despite similar doses used. CONCLUSIONS: Our findings provide guidance for the design of human in vitro CS exposure models in experimental and translational lung research.
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Publication type Article: Journal article
Document type Scientific Article
Keywords Bronchial Epithelial Cells ; Cigarette Smoke ; Dose ; Primary Human Cells ; Validation; Particle Deposition Patterns; Iv Alcohol-dehydrogenase; Induced Oxidative Stress; Human Airway Epithelium; Whole Cigarette-smoke; Gene-expression; Response Element; Hydrocarbon Receptor; Liquid Interface; Up-regulation
Language english
Publication Year 2021
HGF-reported in Year 2021
ISSN (print) / ISBN 1040-0605
e-ISSN 1522-1504
Journal American Journal of Physiology - Lung Cellular and Molecular Physiology
Quellenangaben Volume: 322, Issue: 1, Pages: L129-L148 Article Number: , Supplement: ,
Publisher American Physiological Society
Publishing Place Bethesda, Md. [u.a.]
Reviewing status Peer reviewed
Institute(s) Institute of Lung Health and Immunity (LHI)
Institute of Virology (VIRO)
POF-Topic(s) 80000 - German Center for Lung Research
30202 - Environmental Health
30203 - Molecular Targets and Therapies
Research field(s) Lung Research
Immune Response and Infection
PSP Element(s) G-501800-817
G-501600-001
G-502700-003
G-505000-008
G-552100-001
Grants | Helmholtz Zentrum München (Helmholtz Centre Munich, German Research Center for Environmental Health)
Deutsche Zentrum für Lungenforschung (DZL)
Deutsche Forschungsgemeinschaft (DFG)
DOI 10.1152/ajplung.00091.2021
WOS ID WOS:000743895200010
Scopus ID 85123529313
PubMed ID 34668416
Erfassungsdatum 2021-12-16
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