Comparison of ex vivo and in vitro intestinal cystic fibrosis models to measure CFTR-dependent ion channel activity

J Cyst Fibros. 2018 May;17(3):316-324. doi: 10.1016/j.jcf.2018.02.007. Epub 2018 Mar 13.

Abstract

Background: New functional assays using primary human intestinal adult stem cell cultures can be valuable tools to study epithelial defects in human diseases such as cystic fibrosis.

Methods: CFTR-mediated ion transport was measured in rectal organoid-derived monolayers grown from subjects with various CFTR mutations and compared to donor-matched intestinal current measurements (ICM) in rectal biopsies and forskolin-induced swelling of rectal organoids.

Results: Rectal organoid-derived monolayers were generated within four days. Ion transport measurements of CFTR function using these monolayers correlated with ICM and organoid swelling (r = 0.73 and 0.79 respectively). Culturing the monolayers under differentiation conditions enhanced the detection of mucus-secreting cells and was accompanied by reduced CFTR function.

Conclusions: CFTR-dependent intestinal epithelial ion transport properties can be measured in rectal organoid-derived monolayers of subjects and correlate with donor-matched ICM and rectal organoid swelling.

Keywords: Biopsies; CFTR; Differentiation; Donor-matched; Electrophysiology; Monolayers; Organoids.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cells, Cultured
  • Cystic Fibrosis Transmembrane Conductance Regulator* / genetics
  • Cystic Fibrosis Transmembrane Conductance Regulator* / metabolism
  • Cystic Fibrosis* / genetics
  • Cystic Fibrosis* / metabolism
  • Epithelial Cells / metabolism*
  • Humans
  • Ion Transport / physiology*
  • Models, Biological
  • Mutation
  • Organoids / physiology
  • Rectum

Substances

  • Cystic Fibrosis Transmembrane Conductance Regulator