Optimization of an air-liquid interface

Autor: 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
Rok vydání: 2021
Předmět:
Zdroj: Journal of Aerosol Science
ISSN: 0021-8502
Popis: 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.
Highlights • Calu-3+MDM model shows the epithelial monolayer integrity under prolonged ALI culture. • Calu-3+MDM model has an increased sensitivity in inflammatory responses to LPS aerosol. • Combination of Calu-3+MDM model is a preferred option for long-term ALI exposure to inhaled aerosols.
Databáze: OpenAIRE
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