Document Type
Article
Publication Date
11-20-2023
Original Citation
Cadena D,
Jangra S,
Yurieva M,
Martinek J,
Callender M,
Coxe M,
Choi A,
García-Bernalt Diego J,
Wu T,
Marches F,
Chaussabel D,
García-Sastre A,
Schotsaert M,
Williams A,
Palucka K.
Protocol for establishing primary human lung organoid-derived air-liquid interface cultures from cryopreserved human lung tissue. STAR Protoc. 2023;4(4):102735.
Keywords
JGM
JAX Source
STAR Protoc. 2023;4(4):102735.
ISSN
2666-1667
PMID
37991921
DOI
https://doi.org/10.1016/j.xpro.2023.102735
Grant
This work is partially supported by NIAID grants U19AI142733 (K.P.) and U19AI135972 (A.G.-S.); R01AI141609 (A.W.); R01AI160706 (MS); NIDDK grant R01DK130425 (M.S.); NCI grants P30 CA034196 (K.P.) and U54CA260560 (A.G.-S.); CRIPT (Center for Research on Influenza Pathogenesis and Transmis- sion); and NIAID funded Center of Excellence for Influenza Research and Response (CEIRR, contract number 75N93021C00014) to A.G.-S.
Abstract
Primary human lung organoid-derived air-liquid interface (ALI) cultures serve as a physiologically relevant model to study human airway epithelium in vitro. Here, we present a protocol for establishing these cultures from cryopreserved human lung tissue. We describe steps for lung tissue cryostorage, tissue dissociation, lung epithelial organoid generation, and ALI culture differentiation. We also include quality control steps and technical readouts for monitoring virus response. This protocol demonstrates severe acute respiratory syndrome coronavirus 2 infection in these cultures as an example of their utility. For complete details on the use and execution of this protocol, please refer to Diana Cadena Castaneda et al. (2023).
Comments
This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).