Details
| Original language | English |
|---|---|
| Article number | 2062 |
| Journal | Cells |
| Volume | 10 |
| Issue number | 8 |
| Publication status | Published - 12 Aug 2021 |
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
ASJC Scopus subject areas
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In: Cells, Vol. 10, No. 8, 2062, 12.08.2021.
Research output: Contribution to journal › Article › Research › peer review
}
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 -