Abstract
Epithelial cysts and organoids are multicellular hollow structures formed by correctly polarized epithelial cells. Important in steering these cysts from single cells is the dynamic regulation of extracellular matrix presented ligands, and matrix dynamics. Here, control over the effective ligand concentration is introduced, decoupled from bulk and local mechanical properties, in synthetic dynamic supramolecular hydrogels formed through noncovalent crosslinking of supramolecular fibers. Control over the effective ligand concentration is realized by 1) keeping the ligand concentration constant, but changing the concentration of nonfunctionalized molecules or by 2) varying the ligand concentration, while keeping the concentration of non-functionalized molecules constant. The results show that in 2D, the effective ligand concentration within the supramolecular fibers rather than gel stiffness (from 0.1 to 8 kPa) regulates epithelial polarity. In 3D, increasing the effective ligand concentration from 0.5 × 10 -3 to 2 × 10 -3 m strengthens the effect of increased gel stiffness from 0.1 to 2 kPa, to synergistically yield more correctly polarized cysts. Through integrin manipulation, it is shown that epithelial polarity is regulated by tension-based homeostasis between cells and matrix. The results reveal the effective ligand concentration as influential factor in regulating epithelial polarity and provide insights on engineering of synthetic biomaterials for cell and organoid culture.
| Original language | English |
|---|---|
| Article number | 2300873 |
| Number of pages | 13 |
| Journal | Advanced Materials |
| Volume | 36 |
| Issue number | 43 |
| Early online date | 1 Jun 2023 |
| DOIs | |
| Publication status | Published - Oct 2024 |
Funding
L.R., M.J.H., C.V.C.B., and P.Y.W.D. contributed equally to this work. The authors gratefully acknowledge Joost Wijnakker, Wim de Lau, and Prof. Hans Clevers (Hubrecht Institute) for useful discussions on epithelial polarity and for providing the TS2/16 antibody. The authors thank Lisanne Kuntz for her work on the 3D immunofluorescence staining protocols, Simone de Jong for providing the ROCK-inhibitor Y27632, Johnick van Sprang for his help in the YAP quantification, and Dylan Mostert and prof. Jan de Boer for useful discussions (all at Eindhoven University of Technology). The authors acknowledge the support of the partners of Regenerative Medicine Crossing Borders (RegMed XB), Powered by Health-Holland, Top Sector Life Sciences & Health and the Dutch Ministry of Education, Culture and Science for the Gravitation Programs (024.003.013 and 024.005.020). L.R., M.J.H., C.V.C.B., and P.Y.W.D. contributed equally to this work. The authors gratefully acknowledge Joost Wijnakker, Wim de Lau, and Prof. Hans Clevers (Hubrecht Institute) for useful discussions on epithelial polarity and for providing the TS2/16 antibody. The authors thank Lisanne Kuntz for her work on the 3D immunofluorescence staining protocols, Simone de Jong for providing the ROCK\u2010inhibitor Y27632, Johnick van Sprang for his help in the YAP quantification, and Dylan Mostert and prof. Jan de Boer for useful discussions (all at Eindhoven University of Technology). The authors acknowledge the support of the partners of Regenerative Medicine Crossing Borders (RegMed XB), Powered by Health\u2010Holland, Top Sector Life Sciences & Health and the Dutch Ministry of Education, Culture and Science for the Gravitation Programs (024.003.013 and 024.005.020).
Keywords
- cell–matrix mechanobiology
- effective ligand concentration
- epithelial cell polarity
- supramolecular hydrogels
- synthetic ECM
- Extracellular Matrix/metabolism
- Cell Polarity
- Biocompatible Materials/chemistry
- Epithelial Cells/cytology
- Humans
- Hydrogels/chemistry
- Integrins/metabolism
- Animals
- Madin Darby Canine Kidney Cells
- Dogs
- Ligands