Optimization of an air-liquid interface in vitro cell co-culture model to estimate the hazard of aerosol exposures

Rui-Wen He; Hedwig M Braakhuis; Rob J Vandebriel; Yvonne C M Staal; Eric R Gremmer; Paul H B Fokkens; Claudia Kemp; Jolanda Vermeulen; Remco H S Westerink; Flemming R Cassee

doi:10.1016/j.jaerosci.2020.105703

Optimization of an air-liquid interface in vitro cell co-culture model to estimate the hazard of aerosol exposures

Rui-Wen He
, Hedwig M Braakhuis
, Rob J Vandebriel
, Yvonne C M Staal
, Eric R Gremmer
, Paul H B Fokkens
, Claudia Kemp
, Jolanda Vermeulen
, Remco H S Westerink
, Flemming R Cassee

IRAS OH Toxicology

Center for Infectious Disease Control,National Institute for Public Health and the Environment (RIVM),The Netherlands.

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Inhalation exposure to environmental and occupational aerosol contaminants is associated with many respiratory health problems. To realistically mimic long-term inhalation exposure for toxicity testing, lung epithelial cells need to maintained and exposed under air-liquid interface (ALI) conditions for a prolonged period of time. In addition, to study cellular responses to aerosol particles, lung epithelial cells have to be co-cultured with macrophages. To that aim, we evaluated human bronchial epithelial Calu-3, 16HBE14o- (16HBE), H292, and BEAS-2B cell lines with respect to epithelial morphology, barrier function and cell viability under prolonged ALI culture conditions. Only the Calu-3 cells can retain the monolayer structure and maintain a strong tight junction under long-term ALI culture at least up to 2 weeks. As such, Calu-3 cells were applied as the structural barrier to create co-culture models with human monocyte-derived macrophages (MDMs) and THP-1 derived macrophages (TDMs). Adhesion of macrophages onto the epithelial monolayer was allowed for 4 h with a density of 5 × 104 macrophages/cm2. In comparison to the Calu-3 mono-culture model, Calu-3 + TDM and Calu-3 + MDM co-culture models showed an increased sensitivity in inflammatory responses to lipopolysaccharide (LPS) aerosol at Day 1 of co-culture, with the Calu-3 + MDM model giving a stronger response than Calu-3 + TDM. Therefore, the epithelial monolayer integrity and increased sensitivity make the Calu-3 + MDM co-culture model a preferred option for ALI exposure to inhaled aerosols for toxicity testing.

Original language	English
Article number	105703
Pages (from-to)	1-14
Number of pages	14
Journal	Journal of Aerosol Science
Volume	153
DOIs	https://doi.org/10.1016/j.jaerosci.2020.105703
Publication status	Published - Mar 2021

Bibliographical note

Funding Information:
This work is funded by the EU-project PATROLS (No. 760813) and the Dutch Ministry of Health, Welfare and Sport (project 5.1.2). We thank Marc Bazen, Victoria de Leeuw and Liset de la Fonteyne-Blankestijn from the National Institute for Public Health and the Environment (RIVM) for their valuable assistance with cell culture, cell staining and fluorescence microscopy. We are also grateful to Dr. Barbara Rothen-Rutishauser, Barbara Drasler and Anne Bannuscher from the Adolphe Merkle Institute, University of Fribourg for their kind assistance with the confocal microscopy. The support provided by China Scholarship Council (CSC) during the PhD period of Rui-Wen He in Utrecht University-Institute for Risk Assessment Studies is also acknowledged.

Funding Information:
This work is funded by the EU-project PATROLS (No. 760813 ) and the Dutch Ministry of Health, Welfare and Sport (project 5.1.2). We thank Marc Bazen, Victoria de Leeuw and Liset de la Fonteyne-Blankestijn from the National Institute for Public Health and the Environment (RIVM) for their valuable assistance with cell culture, cell staining and fluorescence microscopy. We are also grateful to Dr. Barbara Rothen-Rutishauser, Barbara Drasler and Anne Bannuscher from the Adolphe Merkle Institute, University of Fribourg for their kind assistance with the confocal microscopy. The support provided by China Scholarship Council (CSC) during the PhD period of Rui-Wen He in Utrecht University-Institute for Risk Assessment Studies is also acknowledged.

Publisher Copyright:
© 2020

Keywords

Aerosol exposures
Air–liquid interface
Barrier function
Co-culture
Epithelial cells
Macrophages

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

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1-s2.0-S0021850220301889-mainFinal published version, 10.1 MBLicence: CC BY

Cite this

He, R.-W., Braakhuis, H. M., Vandebriel, R. J., Staal, Y. C. M., Gremmer, E. R., Fokkens, P. H. B., Kemp, C., Vermeulen, J., Westerink, R. H. S., & Cassee, F. R. (2021). Optimization of an air-liquid interface in vitro cell co-culture model to estimate the hazard of aerosol exposures. Journal of Aerosol Science, 153, 1-14. Article 105703. https://doi.org/10.1016/j.jaerosci.2020.105703

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title = "Optimization of an air-liquid interface in vitro cell co-culture model to estimate the hazard of aerosol exposures",

abstract = "Inhalation exposure to environmental and occupational aerosol contaminants is associated with many respiratory health problems. To realistically mimic long-term inhalation exposure for toxicity testing, lung epithelial cells need to maintained and exposed under air-liquid interface (ALI) conditions for a prolonged period of time. In addition, to study cellular responses to aerosol particles, lung epithelial cells have to be co-cultured with macrophages. To that aim, we evaluated human bronchial epithelial Calu-3, 16HBE14o- (16HBE), H292, and BEAS-2B cell lines with respect to epithelial morphology, barrier function and cell viability under prolonged ALI culture conditions. Only the Calu-3 cells can retain the monolayer structure and maintain a strong tight junction under long-term ALI culture at least up to 2 weeks. As such, Calu-3 cells were applied as the structural barrier to create co-culture models with human monocyte-derived macrophages (MDMs) and THP-1 derived macrophages (TDMs). Adhesion of macrophages onto the epithelial monolayer was allowed for 4 h with a density of 5 × 104 macrophages/cm2. In comparison to the Calu-3 mono-culture model, Calu-3 + TDM and Calu-3 + MDM co-culture models showed an increased sensitivity in inflammatory responses to lipopolysaccharide (LPS) aerosol at Day 1 of co-culture, with the Calu-3 + MDM model giving a stronger response than Calu-3 + TDM. Therefore, the epithelial monolayer integrity and increased sensitivity make the Calu-3 + MDM co-culture model a preferred option for ALI exposure to inhaled aerosols for toxicity testing.",

keywords = "Aerosol exposures, Air–liquid interface, Barrier function, Co-culture, Epithelial cells, Macrophages",

author = "Rui-Wen He and Braakhuis, \{Hedwig M\} and Vandebriel, \{Rob J\} and Staal, \{Yvonne C M\} and Gremmer, \{Eric R\} and Fokkens, \{Paul H B\} and Claudia Kemp and Jolanda Vermeulen and Westerink, \{Remco H S\} and Cassee, \{Flemming R\}",

note = "Funding Information: This work is funded by the EU-project PATROLS (No. 760813) and the Dutch Ministry of Health, Welfare and Sport (project 5.1.2). We thank Marc Bazen, Victoria de Leeuw and Liset de la Fonteyne-Blankestijn from the National Institute for Public Health and the Environment (RIVM) for their valuable assistance with cell culture, cell staining and fluorescence microscopy. We are also grateful to Dr. Barbara Rothen-Rutishauser, Barbara Drasler and Anne Bannuscher from the Adolphe Merkle Institute, University of Fribourg for their kind assistance with the confocal microscopy. The support provided by China Scholarship Council (CSC) during the PhD period of Rui-Wen He in Utrecht University-Institute for Risk Assessment Studies is also acknowledged. Funding Information: This work is funded by the EU-project PATROLS (No. 760813 ) and the Dutch Ministry of Health, Welfare and Sport (project 5.1.2). We thank Marc Bazen, Victoria de Leeuw and Liset de la Fonteyne-Blankestijn from the National Institute for Public Health and the Environment (RIVM) for their valuable assistance with cell culture, cell staining and fluorescence microscopy. We are also grateful to Dr. Barbara Rothen-Rutishauser, Barbara Drasler and Anne Bannuscher from the Adolphe Merkle Institute, University of Fribourg for their kind assistance with the confocal microscopy. The support provided by China Scholarship Council (CSC) during the PhD period of Rui-Wen He in Utrecht University-Institute for Risk Assessment Studies is also acknowledged. Publisher Copyright: {\textcopyright} 2020",

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AU - He, Rui-Wen

AU - Braakhuis, Hedwig M

AU - Vandebriel, Rob J

AU - Staal, Yvonne C M

AU - Gremmer, Eric R

AU - Fokkens, Paul H B

AU - Kemp, Claudia

AU - Vermeulen, Jolanda

AU - Westerink, Remco H S

AU - Cassee, Flemming R

N1 - Funding Information: This work is funded by the EU-project PATROLS (No. 760813) and the Dutch Ministry of Health, Welfare and Sport (project 5.1.2). We thank Marc Bazen, Victoria de Leeuw and Liset de la Fonteyne-Blankestijn from the National Institute for Public Health and the Environment (RIVM) for their valuable assistance with cell culture, cell staining and fluorescence microscopy. We are also grateful to Dr. Barbara Rothen-Rutishauser, Barbara Drasler and Anne Bannuscher from the Adolphe Merkle Institute, University of Fribourg for their kind assistance with the confocal microscopy. The support provided by China Scholarship Council (CSC) during the PhD period of Rui-Wen He in Utrecht University-Institute for Risk Assessment Studies is also acknowledged. Funding Information: This work is funded by the EU-project PATROLS (No. 760813 ) and the Dutch Ministry of Health, Welfare and Sport (project 5.1.2). We thank Marc Bazen, Victoria de Leeuw and Liset de la Fonteyne-Blankestijn from the National Institute for Public Health and the Environment (RIVM) for their valuable assistance with cell culture, cell staining and fluorescence microscopy. We are also grateful to Dr. Barbara Rothen-Rutishauser, Barbara Drasler and Anne Bannuscher from the Adolphe Merkle Institute, University of Fribourg for their kind assistance with the confocal microscopy. The support provided by China Scholarship Council (CSC) during the PhD period of Rui-Wen He in Utrecht University-Institute for Risk Assessment Studies is also acknowledged. Publisher Copyright: © 2020

PY - 2021/3

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N2 - Inhalation exposure to environmental and occupational aerosol contaminants is associated with many respiratory health problems. To realistically mimic long-term inhalation exposure for toxicity testing, lung epithelial cells need to maintained and exposed under air-liquid interface (ALI) conditions for a prolonged period of time. In addition, to study cellular responses to aerosol particles, lung epithelial cells have to be co-cultured with macrophages. To that aim, we evaluated human bronchial epithelial Calu-3, 16HBE14o- (16HBE), H292, and BEAS-2B cell lines with respect to epithelial morphology, barrier function and cell viability under prolonged ALI culture conditions. Only the Calu-3 cells can retain the monolayer structure and maintain a strong tight junction under long-term ALI culture at least up to 2 weeks. As such, Calu-3 cells were applied as the structural barrier to create co-culture models with human monocyte-derived macrophages (MDMs) and THP-1 derived macrophages (TDMs). Adhesion of macrophages onto the epithelial monolayer was allowed for 4 h with a density of 5 × 104 macrophages/cm2. In comparison to the Calu-3 mono-culture model, Calu-3 + TDM and Calu-3 + MDM co-culture models showed an increased sensitivity in inflammatory responses to lipopolysaccharide (LPS) aerosol at Day 1 of co-culture, with the Calu-3 + MDM model giving a stronger response than Calu-3 + TDM. Therefore, the epithelial monolayer integrity and increased sensitivity make the Calu-3 + MDM co-culture model a preferred option for ALI exposure to inhaled aerosols for toxicity testing.

AB - Inhalation exposure to environmental and occupational aerosol contaminants is associated with many respiratory health problems. To realistically mimic long-term inhalation exposure for toxicity testing, lung epithelial cells need to maintained and exposed under air-liquid interface (ALI) conditions for a prolonged period of time. In addition, to study cellular responses to aerosol particles, lung epithelial cells have to be co-cultured with macrophages. To that aim, we evaluated human bronchial epithelial Calu-3, 16HBE14o- (16HBE), H292, and BEAS-2B cell lines with respect to epithelial morphology, barrier function and cell viability under prolonged ALI culture conditions. Only the Calu-3 cells can retain the monolayer structure and maintain a strong tight junction under long-term ALI culture at least up to 2 weeks. As such, Calu-3 cells were applied as the structural barrier to create co-culture models with human monocyte-derived macrophages (MDMs) and THP-1 derived macrophages (TDMs). Adhesion of macrophages onto the epithelial monolayer was allowed for 4 h with a density of 5 × 104 macrophages/cm2. In comparison to the Calu-3 mono-culture model, Calu-3 + TDM and Calu-3 + MDM co-culture models showed an increased sensitivity in inflammatory responses to lipopolysaccharide (LPS) aerosol at Day 1 of co-culture, with the Calu-3 + MDM model giving a stronger response than Calu-3 + TDM. Therefore, the epithelial monolayer integrity and increased sensitivity make the Calu-3 + MDM co-culture model a preferred option for ALI exposure to inhaled aerosols for toxicity testing.

KW - Aerosol exposures

KW - Air–liquid interface

KW - Barrier function

KW - Co-culture

KW - Epithelial cells

KW - Macrophages

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M1 - 105703

ER -

Optimization of an air-liquid interface in vitro cell co-culture model to estimate the hazard of aerosol exposures

Abstract

Bibliographical note

Keywords

UN SDGs

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