Physiological shear stress enhances differentiation, mucus-formation and structural 3D organization of intestinal epithelial cells in vitro

verfasst von: Marcus Lindner, Anna Laporte, Stephan Block, Laura Elomaa, Marie Weinhart
Abstract: Gastrointestinal (GI) mucus plays a pivotal role in the tissue homoeostasis and function-ality of the gut. However, due to the shortage of affordable, realistic in vitro GI models with a physiologically relevant mucus layer, studies with deeper insights into structural and compositional changes upon chemical or physical manipulation of the system are rare. To obtain an improved mucus-containing cell model, we developed easy-to-use, reusable culture chambers that facilitated the application of GI shear stresses (0.002–0.08 dyn·cm⁻² ) to cells on solid surfaces or membranes of cell culture inserts in bioreactor systems, thus making them readily accessible for subsequent analyses, e.g., by confocal microscopy or transepithelial electrical resistance (TEER) measurement. The human mucus-producing epithelial HT29-MTX cell-line exhibited superior reorganization into 3-dimensional villi-like structures with highly proliferative tips under dynamic culture conditions when compared to static culture (up to 180 vs. 80 µm in height). Additionally, the median mucus layer thickness was significantly increased under flow (50 ± 24 vs. 29 ± 14 µm (static)), with a simultaneous accelerated maturation of the cells into a goblet-like phenotype. We demonstrated the strong impact of culture conditions on the differentiation and reorganization of HT29-MTX cells. The results comprise valuable advances towards the improvement of existing GI and mucus models or the development of novel systems using our newly designed culture chambers.
Organisationseinheit(en): Institut für Physikalische Chemie und Elektrochemie
Externe Organisation(en): Freie Universität Berlin (FU Berlin)
Typ: Artikel
Journal: Cells
Band: 10
ISSN: 2073-4409
Publikationsdatum: 12.08.2021
Publikationsstatus: Veröffentlicht
Peer-reviewed: Ja
ASJC Scopus Sachgebiete: Allgemeine Medizin
Elektronische Version(en): https://doi.org/10.3390/cells10082062 (Zugang: Offen)

BibTeX

@article{946925c7e49e4884a708f28ace3c3025,
title = "Physiological shear stress enhances differentiation, mucus-formation and structural 3D organization of intestinal epithelial cells in vitro",
abstract = "Gastrointestinal (GI) mucus plays a pivotal role in the tissue homoeostasis and function-ality of the gut. However, due to the shortage of affordable, realistic in vitro GI models with a physiologically relevant mucus layer, studies with deeper insights into structural and compositional changes upon chemical or physical manipulation of the system are rare. To obtain an improved mucus-containing cell model, we developed easy-to-use, reusable culture chambers that facilitated the application of GI shear stresses (0.002–0.08 dyn·cm−2 ) to cells on solid surfaces or membranes of cell culture inserts in bioreactor systems, thus making them readily accessible for subsequent analyses, e.g., by confocal microscopy or transepithelial electrical resistance (TEER) measurement. The human mucus-producing epithelial HT29-MTX cell-line exhibited superior reorganization into 3-dimensional villi-like structures with highly proliferative tips under dynamic culture conditions when compared to static culture (up to 180 vs. 80 µm in height). Additionally, the median mucus layer thickness was significantly increased under flow (50 ± 24 vs. 29 ± 14 µm (static)), with a simultaneous accelerated maturation of the cells into a goblet-like phenotype. We demonstrated the strong impact of culture conditions on the differentiation and reorganization of HT29-MTX cells. The results comprise valuable advances towards the improvement of existing GI and mucus models or the development of novel systems using our newly designed culture chambers.",
keywords = "3D-printed insert chamber, Bioreactor, Cell-based mucus model, Cellular self-organization, CFD simulation, Goblet cell differentiation, Native mucus thickness, Physiological fluid flow, Reverse cell culture",
author = "Marcus Lindner and Anna Laporte and Stephan Block and Laura Elomaa and Marie Weinhart",
year = "2021",
month = aug,
day = "12",
doi = "10.3390/cells10082062",
language = "English",
volume = "10",
journal = "Cells",
issn = "2073-4409",
publisher = "MDPI Multidisciplinary Digital Publishing Institute",
number = "8",
}