์ฝ˜ํ…์ธ ๋กœ ๊ฑด๋„ˆ๋›ฐ๊ธฐ
Merck

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
์ดˆ๋ก

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
์ œํ’ˆ ๋ฒˆํ˜ธ
๋ธŒ๋žœ๋“œ
์ œํ’ˆ ์„ค๋ช…

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