TY - JOUR

T1 - Physiological shear stress enhances differentiation, mucus-formation and structural 3D organization of intestinal epithelial cells in vitro

AU - Lindner, Marcus

AU - Laporte, Anna

AU - Block, Stephan

AU - Elomaa, Laura

AU - Weinhart, Marie

PY - 2021/8/12

Y1 - 2021/8/12

N2 - 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.

AB - 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.

KW - 3D-printed insert chamber

KW - Bioreactor

KW - Cell-based mucus model

KW - Cellular self-organization

KW - CFD simulation

KW - Goblet cell differentiation

KW - Native mucus thickness

KW - Physiological fluid flow

KW - Reverse cell culture

UR - http://www.scopus.com/inward/record.url?scp=85115044846&partnerID=8YFLogxK

U2 - 10.3390/cells10082062

DO - 10.3390/cells10082062

M3 - Article

C2 - 34440830

AN - SCOPUS:85115044846

VL - 10

JO - Cells

JF - Cells

SN - 2073-4409

IS - 8

M1 - 2062

ER -