Institute for Complex Molecular Systems, Eindhoven University of Technology, PO Box 513, Eindhoven, The Netherlands.
Department of Biomedical Engineering, Laboratory of Chemical Biology, Eindhoven University of Technology, PO Box 513, Eindhoven, The Netherlands.
Adv Mater. 2024 Oct;36(43):e2300873. doi: 10.1002/adma.202300873. Epub 2023 Jul 20.
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 to 2 × 10 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.
上皮细胞囊泡和类器官是由极性正确的上皮细胞形成的多细胞中空结构。重要的是，通过对细胞外基质配体的动态调控以及基质动力学，将这些细胞从单细胞状态引导至上皮细胞囊泡和类器官状态。在这里，通过超分子纤维的非共价交联形成的合成动态超分子水凝胶中，实现了对有效配体浓度的控制，使其与体相和局部力学性能解耦。通过以下两种方式实现了对有效配体浓度的控制：1）保持配体浓度恒定，改变非功能化分子的浓度；或 2）改变配体浓度，同时保持非功能化分子的浓度恒定。结果表明，在 2D 中，超分子纤维内的有效配体浓度（从 0.1 到 8 kPa）而不是凝胶刚度调节上皮细胞极性。在 3D 中，将有效配体浓度从 0.5×10-6 增加到 2×10-6 m，可增强凝胶刚度从 0.1 到 2 kPa 的增加对上皮细胞极性的影响，从而协同产生更多极性正确的囊泡。通过整合素操作，表明上皮细胞极性受细胞与基质之间基于张力的平衡的调节。结果揭示了有效配体浓度作为调节上皮细胞极性的影响因素，并为用于细胞和类器官培养的合成生物材料的工程设计提供了新的见解。
