Permeability analyses and three dimensional imaging of interferon gamma-induced barrier disintegration in intestinal organoids

Journal article


Publication Details

Author(s): Bardenbacher M, Ruder B, Britzen-Laurent N, Schmid B, Waldner M, Naschberger E, Scharl M, Mueller W, Günther C, Becker C, Stürzl M, Tripal P
Journal: Stem Cell Research
Publication year: 2019
Volume: 35
ISSN: 1873-5061


Abstract

The aberrant regulation of the epithelial barrier integrity is involved in many diseases of the digestive tract, including inflammatory bowel diseases and colorectal cancer. Intestinal epithelial cell organoid cultures provide new perspectives for analyses of the intestinal barrier in vitro. However, established methods of barrier function analyses from two dimensional cultures have to be adjusted to the analysis of three dimensional organoid structures. Here we describe the methodology for analysis of epithelial barrier function and molecular regulation in intestinal organoids. Barrier responses to interferon-y of intestinal organoids with and without epithelial cellspecific deletion of the interferon-y-receptor 2 gene were used as a model system. The established method allowed monitoring of the kinetics of interferon-y-induced permeability changes in living organoids. Proteolytic degradation and altered localization of the tight junction proteins claudin-2, -7, and - 15 was detected using confocal spinning disc microscopy with 3D reconstruction. Hessian analysis was used for quantification of relocalization of claudins. In summary, we provide a novel methodologic approach for quantitative analyses of intestinal epithelial barrier functions in the 3D organoid model.


FAU Authors / FAU Editors

Becker, Christoph Prof. Dr. rer. nat.
Professur für Molekulare Gastroenterologie
Medizinische Klinik 1 - Gastroenterologie, Pneumologie und Endokrinologie
Naschberger, Elisabeth PD Dr.
Medizinische Fakultät
Schmid, Benjamin Dr.
Optical Imaging Center Erlangen (OICE)
Stürzl, Michael Prof. Dr.
Professur für Molekulare und Experimentelle Chirurgie
Tripal, Philipp Dr.
Chirurgische Klinik
Waldner, Maximilian Prof. Dr.
Lehrstuhl für Innere Medizin I


External institutions with authors

Universitätsspital Zürich (USZ)
University of Manchester


How to cite

APA:
Bardenbacher, M., Ruder, B., Britzen-Laurent, N., Schmid, B., Waldner, M., Naschberger, E.,... Tripal, P. (2019). Permeability analyses and three dimensional imaging of interferon gamma-induced barrier disintegration in intestinal organoids. Stem Cell Research, 35. https://dx.doi.org/10.1016/j.scr.2019.101383

MLA:
Bardenbacher, Marco, et al. "Permeability analyses and three dimensional imaging of interferon gamma-induced barrier disintegration in intestinal organoids." Stem Cell Research 35 (2019).

BibTeX: 

Last updated on 2019-20-03 at 15:38