University of Mons, Laboratory for Complex Fluids and Interfaces, Mechanobiology and Soft Matter Group, CIRMAP, Research Institute for Biosciences, Place du Parc, Mons, Belgium.
Biotechnol Bioeng. 2020 Sep;117(9):2887-2896. doi: 10.1002/bit.27446. Epub 2020 Jun 16.
The development of epithelial lumens in ducts is essential to the functioning of various organs and in organogenesis. Ductal elongation requires the collective migration of cell cohorts in three-dimensional (3D) confined spaces, while maintaining their epithelial integrity. Epithelial lumens generally adopt circular morphologies, however abnormalities in complex physiological environments can lead to the narrowing of glandular spaces that adopt elongated and slit-like morphologies. Here, we describe a simple method to form epithelial tissues in microchannels of various widths (100-300 µm) with a constant height of 25 µm that mimic elongated geometries of glandular spaces. The significance of this biomimetic platform has been evidenced by studying the migration of epithelial cell sheets inside these narrow slits of varying dimensions. We show that the growth of epithelial tissues in 3D-confined slits leads to a gradient of cell density along the slit axis and that the migration cell velocity depends on the extent of the spatial confinement. Our findings indicate that nuclear orientation is higher for leader cells and depends on the slit width, whereas YAP protein was predominantly localized in the nucleus of leader cells. This method will pave the way to studies aiming at understanding how 3D-confined spaces, which are reminiscent of in vivo pathological conditions, can affect the growth and the homeostasis of epithelial tissues.
导管中上皮腔的发育对于各种器官的功能和器官发生至关重要。导管的伸长需要细胞群体在三维（3D）受限空间中的集体迁移，同时保持其上皮完整性。上皮腔通常采用圆形形态，然而在复杂的生理环境中出现异常会导致腺腔变窄，呈现出拉长的缝隙样形态。在这里，我们描述了一种简单的方法，可在高度为 25μm、宽度为 100-300μm 的各种微通道中形成具有上皮组织，这些微通道的形状模仿了腺腔的伸长几何形状。通过研究上皮细胞片在这些具有不同尺寸的狭窄缝隙中的迁移，证明了这种仿生平台的重要意义。我们表明，在 3D 受限缝隙中生长的上皮组织会导致细胞密度沿缝隙轴呈梯度分布，并且迁移细胞的速度取决于空间限制的程度。我们的发现表明，对于先导细胞，核取向更高，并取决于缝隙的宽度，而 YAP 蛋白主要定位于先导细胞的核内。这种方法将为旨在了解类似于体内病理条件的 3D 受限空间如何影响上皮组织的生长和内稳态的研究铺平道路。
