Approaches to Study Differentiation and Repair of Human Airway Epithelial Cells

Crespin, Sophie; Bacchetta, Marc; Huang, Song; Dudez, Tecla; Wiszniewski, Ludovic; Chanson, Marc

doi:10.1007/978-1-61779-120-8_10

Cited by 11 publications

(8 citation statements)

References 14 publications

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“…The basal medium, which consisted of DMEM:F12 (3:1, Life Technologies, Zug, Switzerland) supplemented with 1.5% Ultroser G (Bioserpa, Cergy, France) and antibiotics, was refreshed every 2 days. Mechanical wounding was performed using an airbrush linked to a pressure regulator, as previously described 15 .…”

Section: Methodsmentioning

confidence: 99%

Transcriptomic profile of cystic fibrosis airway epithelial cells undergoing repair

et al. 2019

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Pathological remodeling of the airway epithelium is commonly observed in Cystic Fibrosis (CF). The different cell types that constitute the airway epithelium are regenerated upon injury to restore integrity and maintenance of the epithelium barrier function. The molecular signature of tissue repair in CF airway epithelial cells has, however, not well been investigated in primary cultures. We therefore collected RNA-seq data from well-differentiated primary cultures of bronchial human airway epithelial cells (HAECs) of CF (F508del/F508del) and non-CF (NCF) origins before and after mechanical wounding, exposed or not to flagellin. We identified the expression changes with time of repair of genes, the products of which are markers of the different cell types that constitute the airway epithelium (basal, suprabasal, intermediate, secretory, goblet and ciliated cells as well as ionocytes). Researchers in the CF field may benefit from this transcriptomic profile, which covers the initial steps of wound repair and revealed differences in this process between CF and NCF cultures.

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Section: Methodsmentioning

confidence: 99%

Transcriptomic profile of cystic fibrosis airway epithelial cells undergoing repair

et al. 2019

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“…There are currently different commercially available respiratory epithelia cultures emulated from the original HBECs model described by Gray and colleagues, notably EpiAirway™ from MatTek and MucilAir™ from Epithelix. These systems form a polarized columnar epithelia when maintained at the air liquid interface (ALI) on transwell porous membranes, in which basal, mucus producing, and ciliated cells can be distinguished (Chemuturi et al, 2005;Crespin et al, 2011;Gray et al, 1996).…”

Section: Introductionmentioning

confidence: 99%

Targeted omics analyses, and metabolic enzyme activity assays demonstrate maintenance of key mucociliary characteristics in long term cultures of reconstituted human airway epithelia

Baxter

Thain²,

Banerjee

et al. 2015

Toxicology in Vitro

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3D reconstituted respiratory epithelia have emerged as better in vitro models for toxicological testing compared to cell lines due to the conservation of key morphological features and functions. MucilAir™ is a commercially available human airway epithelia system that can potentially maintain functional attributes for up to a year, however, detailed mucociliary characteristics and xenobiotic metabolism relevant to inhaled pro-toxicant bioactivation is lacking. Here, we assessed in MucilAir™ some key biomarkers that are characteristic of the respiratory epithelia including morphology, function and xenobiotics metabolism. The end points that were measured included targeted proteomics using a panel of 243 airway surface liquid (ASL) proteins, cilia beat frequency (CBF), a qRT-PCR screen of xenobiotic metabolizing enzymes, and CYP2A6/13, CYP1A1/1B1 activity. Comparison of ASL proteomics with human sputum identified key proteins common to both matrices, but present at different levels. Xenobiotic metabolism gene profiling demonstrated strong similarities with the normal human lung and did not reveal any consistent changes when assessed over a 6 month period. Inducibility and activity of CYP1A1/1B1 and activity of CYP2A6/2A13 were present at one month in culture and maintained in one tested MucilAir™ donor for several months. In conclusion, MucilAir™ presented important morphological and metabolic characteristics of a mucociliary epithelium in short and long term culture. MucilAir™ is therefore a potentially useful model to test repeated sub-cytotoxic doses of toxicants.

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“…Pharmacological correction of CFTR function in primary nasal HAECs can be used as a biomarker in personalized CF treatment. These cultures, which can regenerate after injury, also represent an excellent model to investigate the mechanisms of differentiation and repair in vitro (50, 51). The CRISPR-Cas9 system has already been applied to both model systems to efficiently knockdown CFTR (52, 53).…”

Section: In Vitro Models To Study Cftr Function and Cf Pathogenesismentioning

confidence: 99%

Animal and model systems for studying cystic fibrosis

Rosen

Chanson

Gawenis

et al. 2018

Journal of Cystic Fibrosis

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The cystic fibrosis (CF) field is the beneficiary of five species of animal models that lack functional cystic fibrosis transmembrane conductance regulator (CFTR) channel. These models are rapidly informing mechanisms of disease pathogenesis and CFTR function regardless of how faithfully a given organ reproduces the human CF phenotype. New approaches of genetic engineering with RNA-guided nucleases are rapidly expanding both the potential types of models available and the approaches to correct the CFTR defect. The application of new CRISPR/Cas9 genome editing techniques are similarly increasing capabilities for in vitro modeling of CFTR functions in cell lines and primary cells using air-liquid interface cultures and organoids. Gene editing of CFTR mutations in somatic stem cells and induced pluripotent stem cells is also transforming gene therapy approaches for CF. This short review evaluates several areas that are key to building animal and cell systems capable of modeling CF disease and testing potential treatments.

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Approaches to Study Differentiation and Repair of Human Airway Epithelial Cells

Cited by 11 publications

References 14 publications

Transcriptomic profile of cystic fibrosis airway epithelial cells undergoing repair

Transcriptomic profile of cystic fibrosis airway epithelial cells undergoing repair

Targeted omics analyses, and metabolic enzyme activity assays demonstrate maintenance of key mucociliary characteristics in long term cultures of reconstituted human airway epithelia

Animal and model systems for studying cystic fibrosis

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