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Article

Physiological Shear Stress Enhances Differentiation, Mucus-Formation and Structural 3D Organization of Intestinal Epithelial Cells In Vitro

1
Institute of Chemistry and Biochemistry, Freie Universität Berlin, 14195 Berlin, Germany
2
Institute of Physical Chemistry and Electrochemistry, Leibniz Universität Hannover, 30167 Hannover, Germany
*
Author to whom correspondence should be addressed.
Academic Editor: Alexander E. Kalyuzhny
Cells 2021, 10(8), 2062; https://doi.org/10.3390/cells10082062
Received: 15 July 2021 / Revised: 6 August 2021 / Accepted: 10 August 2021 / Published: 12 August 2021
(This article belongs to the Section Cell Methods)
Gastrointestinal (GI) mucus plays a pivotal role in the tissue homoeostasis and functionality 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. View Full-Text
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 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
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MDPI and ACS Style

Lindner, M.; Laporte, A.; Block, S.; Elomaa, L.; Weinhart, M. Physiological Shear Stress Enhances Differentiation, Mucus-Formation and Structural 3D Organization of Intestinal Epithelial Cells In Vitro. Cells 2021, 10, 2062. https://doi.org/10.3390/cells10082062

AMA Style

Lindner M, Laporte A, Block S, Elomaa L, Weinhart M. Physiological Shear Stress Enhances Differentiation, Mucus-Formation and Structural 3D Organization of Intestinal Epithelial Cells In Vitro. Cells. 2021; 10(8):2062. https://doi.org/10.3390/cells10082062

Chicago/Turabian Style

Lindner, Marcus, Anna Laporte, Stephan Block, Laura Elomaa, and Marie Weinhart. 2021. "Physiological Shear Stress Enhances Differentiation, Mucus-Formation and Structural 3D Organization of Intestinal Epithelial Cells In Vitro" Cells 10, no. 8: 2062. https://doi.org/10.3390/cells10082062

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