Isolation, cultivation, and application of primary respiratory epithelial cells obtained by nasal brushing, polyp samples, or lung explants

Anita Golec; Iwona  Pranke; Paolo  Scuderi; Kate Hayes; Elise  Dreano; Fiona Dunlevy; Aurelie Hatton; Damian G. Downey; Luis Galietta; Isabelle  Sermet

doi:10.1016/j.xpro.2022.101419

Isolation, cultivation, and application of primary respiratory epithelial cells obtained by nasal brushing, polyp samples, or lung explants

Anita Golec, Iwona Pranke, Paolo Scuderi, Kate Hayes, Elise Dreano, Fiona Dunlevy, Aurelie Hatton, Damian G. Downey, Luis Galietta, Isabelle Sermet

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

6 Citations (Scopus)

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Abstract

Here, we present a standardized protocol for isolation, maintenance, and polarization of the respiratory epithelial primary cells from patient samples acquired from nasal brushing, polyp specimens, or lung explants. This protocol generates a clearly defined polarized layer of epithelial cells on filters, with a good number of ciliated cells and a thin layer of mucus. We detail the steps for samples prepared from patients with cystic fibrosis as well as from subjects without cystic fibrosis.

Original language	English
Article number	101419
Number of pages	33
Journal	STAR Protocols
Volume	3
Issue number	2
Early online date	27 May 2022
DOIs	https://doi.org/10.1016/j.xpro.2022.101419
Publication status	Published - 17 Jun 2022

Bibliographical note

Funding Information:
We would like to express our gratitude to Prof. Jean-François Papon, Ciliary Dyskinesia Centre, Bicêtre Hôpital, Paris, France; Dr. Charlotte Roy, Hôpital Necker Enfants Malades, Paris, France; and Dr. Caroline Faucon, Centre Hospitalier Universitaire, Caen, France for assistance with video production and discussions regarding practical aspects of the protocol. We would also like to express our gratitude to the laboratory staff at INSERM, Paris, France for helpful remarks. ECFS-CTN is a partner in the European Reference Network: ERN-LUNG. All authors contributed to the design, draft of the protocol, and final review of the manuscript. Damian Downey has received consultancy fees and/or grant support from Vertex pharmaceuticals, Proteostasis Therapeutics, Gilead, Chiesi, and Insmed. He is Director of the European CF Society – Clinical Trials Network and a member of the UK CF Trust Research Scientific Oversight Board of the Clinical Trials Accelerator Platform. Isabelle Sermet has received consultancy fees and/or grant support from Vertex Therapeutics, Eloxx, and PTC Therapeutics and is a Board Member of the European Cystic Fibrosis Society. Luis Galietta is a member of the Advisory Board of Enterprise Therapeutics.

Funding Information:
Damian Downey has received consultancy fees and/or grant support from Vertex pharmaceuticals, Proteostasis Therapeutics, Gilead, Chiesi, and Insmed. He is Director of the European CF Society – Clinical Trials Network and a member of the UK CF Trust Research Scientific Oversight Board of the Clinical Trials Accelerator Platform.

Funding Information:
Isabelle Sermet has received consultancy fees and/or grant support from Vertex Therapeutics, Eloxx, and PTC Therapeutics and is a Board Member of the European Cystic Fibrosis Society.

Publisher Copyright:
© 2022 The Author(s)

Keywords

Nasal epithelium
respiratory diseases
epithelial cells

ASJC Scopus subject areas

General Biochemistry,Genetics and Molecular Biology
General Neuroscience
General Immunology and Microbiology
General Medicine

UN SDGs

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

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10.1016/j.xpro.2022.101419Licence: CC BY

Isolation, cultivation, and application of primary respiratory epithelial cells obtained by nasal brushing, polyp samples, or lung explants
Copyright 2022 the authors. This is an open access article published under a Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution and reproduction in any medium, provided the author and source are cited.
Final published version, 3.81 MBLicence: CC BY

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Cite this

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title = "Isolation, cultivation, and application of primary respiratory epithelial cells obtained by nasal brushing, polyp samples, or lung explants",

abstract = "Here, we present a standardized protocol for isolation, maintenance, and polarization of the respiratory epithelial primary cells from patient samples acquired from nasal brushing, polyp specimens, or lung explants. This protocol generates a clearly defined polarized layer of epithelial cells on filters, with a good number of ciliated cells and a thin layer of mucus. We detail the steps for samples prepared from patients with cystic fibrosis as well as from subjects without cystic fibrosis.",

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note = "Funding Information: We would like to express our gratitude to Prof. Jean-Fran{\c c}ois Papon, Ciliary Dyskinesia Centre, Bic{\^e}tre H{\^o}pital, Paris, France; Dr. Charlotte Roy, H{\^o}pital Necker Enfants Malades, Paris, France; and Dr. Caroline Faucon, Centre Hospitalier Universitaire, Caen, France for assistance with video production and discussions regarding practical aspects of the protocol. We would also like to express our gratitude to the laboratory staff at INSERM, Paris, France for helpful remarks. ECFS-CTN is a partner in the European Reference Network: ERN-LUNG. All authors contributed to the design, draft of the protocol, and final review of the manuscript. Damian Downey has received consultancy fees and/or grant support from Vertex pharmaceuticals, Proteostasis Therapeutics, Gilead, Chiesi, and Insmed. He is Director of the European CF Society – Clinical Trials Network and a member of the UK CF Trust Research Scientific Oversight Board of the Clinical Trials Accelerator Platform. Isabelle Sermet has received consultancy fees and/or grant support from Vertex Therapeutics, Eloxx, and PTC Therapeutics and is a Board Member of the European Cystic Fibrosis Society. Luis Galietta is a member of the Advisory Board of Enterprise Therapeutics. Funding Information: Damian Downey has received consultancy fees and/or grant support from Vertex pharmaceuticals, Proteostasis Therapeutics, Gilead, Chiesi, and Insmed. He is Director of the European CF Society – Clinical Trials Network and a member of the UK CF Trust Research Scientific Oversight Board of the Clinical Trials Accelerator Platform. Funding Information: Isabelle Sermet has received consultancy fees and/or grant support from Vertex Therapeutics, Eloxx, and PTC Therapeutics and is a Board Member of the European Cystic Fibrosis Society. Publisher Copyright: {\textcopyright} 2022 The Author(s)",

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AU - Golec, Anita

AU - Pranke, Iwona

AU - Scuderi, Paolo

AU - Hayes, Kate

AU - Dreano, Elise

AU - Dunlevy, Fiona

AU - Hatton, Aurelie

AU - Downey, Damian G.

AU - Galietta, Luis

AU - Sermet, Isabelle

N1 - Funding Information: We would like to express our gratitude to Prof. Jean-François Papon, Ciliary Dyskinesia Centre, Bicêtre Hôpital, Paris, France; Dr. Charlotte Roy, Hôpital Necker Enfants Malades, Paris, France; and Dr. Caroline Faucon, Centre Hospitalier Universitaire, Caen, France for assistance with video production and discussions regarding practical aspects of the protocol. We would also like to express our gratitude to the laboratory staff at INSERM, Paris, France for helpful remarks. ECFS-CTN is a partner in the European Reference Network: ERN-LUNG. All authors contributed to the design, draft of the protocol, and final review of the manuscript. Damian Downey has received consultancy fees and/or grant support from Vertex pharmaceuticals, Proteostasis Therapeutics, Gilead, Chiesi, and Insmed. He is Director of the European CF Society – Clinical Trials Network and a member of the UK CF Trust Research Scientific Oversight Board of the Clinical Trials Accelerator Platform. Isabelle Sermet has received consultancy fees and/or grant support from Vertex Therapeutics, Eloxx, and PTC Therapeutics and is a Board Member of the European Cystic Fibrosis Society. Luis Galietta is a member of the Advisory Board of Enterprise Therapeutics. Funding Information: Damian Downey has received consultancy fees and/or grant support from Vertex pharmaceuticals, Proteostasis Therapeutics, Gilead, Chiesi, and Insmed. He is Director of the European CF Society – Clinical Trials Network and a member of the UK CF Trust Research Scientific Oversight Board of the Clinical Trials Accelerator Platform. Funding Information: Isabelle Sermet has received consultancy fees and/or grant support from Vertex Therapeutics, Eloxx, and PTC Therapeutics and is a Board Member of the European Cystic Fibrosis Society. Publisher Copyright: © 2022 The Author(s)

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N2 - Here, we present a standardized protocol for isolation, maintenance, and polarization of the respiratory epithelial primary cells from patient samples acquired from nasal brushing, polyp specimens, or lung explants. This protocol generates a clearly defined polarized layer of epithelial cells on filters, with a good number of ciliated cells and a thin layer of mucus. We detail the steps for samples prepared from patients with cystic fibrosis as well as from subjects without cystic fibrosis.

AB - Here, we present a standardized protocol for isolation, maintenance, and polarization of the respiratory epithelial primary cells from patient samples acquired from nasal brushing, polyp specimens, or lung explants. This protocol generates a clearly defined polarized layer of epithelial cells on filters, with a good number of ciliated cells and a thin layer of mucus. We detail the steps for samples prepared from patients with cystic fibrosis as well as from subjects without cystic fibrosis.

KW - Nasal epithelium

KW - respiratory diseases

KW - epithelial cells

U2 - 10.1016/j.xpro.2022.101419

DO - 10.1016/j.xpro.2022.101419

M3 - Article

C2 - 35664255

AN - SCOPUS:85131069652

SN - 2666-1667

VL - 3

JO - STAR Protocols

JF - STAR Protocols

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M1 - 101419

ER -

Isolation, cultivation, and application of primary respiratory epithelial cells obtained by nasal brushing, polyp samples, or lung explants

Abstract

Bibliographical note

Keywords

ASJC Scopus subject areas

UN SDGs

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