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

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

STAR protocols (2022-06-07)
Anita Golec, Iwona Pranke, Paolo Scudieri, Kate Hayes, Elise Dreano, Fiona Dunlevy, Aurelie Hatton, Damian G Downey, Luis Galietta, Isabelle Sermet
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.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
HEPES, ≥99.5% (titration)
Sigma-Aldrich
(−)-Epinephrine (+)-bitartrate salt, solid
Sigma-Aldrich
Hydrocortisone solution, 50 μM, sterile-filtered, BioXtra, suitable for cell culture
Sigma-Aldrich
Collagen from human placenta, Bornstein and Traub Type IV, powder