Aerosol-Cell Exposure System Applied to Semi-Adherent Cells for Aerosolization of Lung Surfactant and Nanoparticles Followed by High Quality RNA Extraction

Overview

Journal Nanomaterials (Basel)

Date 2022 Apr 23

PMID 35458071

Authors

Melanie M Leroux

Romain Hocquel

Kevin Bourge

Bostjan Kokot

Hana Kokot

Tilen Koklic

Janez Strancar

Yaobo Ding

Pramod Kumar

Otmar Schmid

Bertrand H Rihn

Luc Ferrari

Olivier Joubert

Affiliations

Soon will be listed here.

Abstract

Objectives: In this work, we developed a comprehensive methodology for coating semi-adherent lung cells cultured at the ALI with aerosolized surfactant and subsequent dose-controlled exposure to nanoparticles (NPs). This protocol is optimized for subsequent transcriptomic studies.

Methods: Semi-adherent rat alveolar macrophages NR8383 were grown at the ALI and coated with lung surfactant through nebulization using the VITROCELL Cloud 6 System before being exposed to TiO NM105 NPs. After NP exposures, RNA was extracted and its quantity and quality were measured.

Results: The VITROCELL Cloud system allowed for uniform and ultrathin coating of cells with aerosolized surfactant mimicking physiological conditions in the lung. While nebulization of 57 μL of 30 mg/mL TiO and 114 μL of 15 mg/mL TiO nanoparticles yielded identical cell delivered dose, the reproducibility of dose as well as the quality of RNA extracted were better for 114 μL.

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