4.3 Article

In vitro 3D culture lung model from expanded primary cystic fibrosis human airway cells

Journal

JOURNAL OF CYSTIC FIBROSIS
Volume 19, Issue 5, Pages 752-761

Publisher

ELSEVIER
DOI: 10.1016/j.jcf.2020.05.007

Keywords

Cystic fibrosis; Lung model; In vitro; CFTR ion channel; Cell expansion

Funding

  1. National Institutes of Health [R01AI124121, R01HL127651]
  2. Cystic Fibrosis Foundation [MCCOY17R2, MCCOY19R0]

Ask authors/readers for more resources

Background: In vitro cystic fibrosis (CF) models are crucial for understanding the mechanisms and consequences of the disease. They are also the gold standard for pre-clinical efficacy studies of current and novel CF drugs. However, few studies have investigated expansion and differentiation of primary CF human bronchial epithelial (CF-HBE) cells. Here we describe culture conditions to expand primary CF airway cells while preserving their ability to differentiate into 3D epithelial cultures expressing functional cystic fibrosis transmembrane conductance regulator (CFTR) ion channels that responds to CFTR modulators. Methods: Primary CF airway cells were expanded using PneumaCult (TM)-Ex Plus (StemCell Technologies) medium with no feeder cells or added Rho kinase (ROCK) inhibitor. Differentially passaged CF-HBE cells at the air-liquid interface (ALI) were characterized phenotypically and functionally in response to the CFTR corrector drug VX-661 (Tezacaftor). Results: CF-HBE primary cells, expanded up to six passages (similar to 25 population doublings), differentiated into 3D epithelial cultures as evidenced by trans-epithelial electrical resistance (TEER) of >400 Ohms.cm(2) and presence of pseudostratified columnar ciliated epithelium with goblet cells. However, up to passage five cells from most donors showed increased CFTR-mediated short-circuit currents when treated with the corrector drug, VX-661. Ciliary beat frequency (CBF) also increased with the corrector VX-661. Conclusions: CF donor-derived airway cells can be expanded without the use of feeder cells or additional ROCK inhibitor, and still achieve optimal 3D epithelial cultures that respond to CFTR modulators. The study of rare CF mutations could benefit from cell expansion and could lead to the design of personalized medicine/treatments. (C) 2020 European Cystic Fibrosis Society. Published by Elsevier B.V. All rights reserved.

Authors

I am an author on this paper
Click your name to claim this paper and add it to your profile.

Reviews

Primary Rating

4.3
Not enough ratings

Secondary Ratings

Novelty
-
Significance
-
Scientific rigor
-
Rate this paper

Recommended

No Data Available
No Data Available