Establishment of a novel in vitro model of stratified epithelial wound healing with barrier function.
Establishment of a novel in vitro model of stratified epithelial wound healing with barrier function.
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DOI：
10.1038/srep19395
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发表时间：
2016-01-13
影响因子：
4.6
通讯作者：
Argüeso P
中科院分区：
文献类型：
--
作者：
Gonzalez-Andrades M;Alonso-Pastor L;Mauris J;Cruzat A;Dohlman CH;Argüeso P
通过上皮细胞的集体运动修复伤口是多细胞生物的基本过程。在角膜和皮肤等复层上皮中，愈合分三个步骤进行，包括潜伏期、迁移期和重建期。已经开发了几种简单且廉价的测定法来研究体外细胞迁移的生物学。然而，这些测定主要基于单层系统，无法重现与多层系统相关的分化过程。在这里，我们描述了一种简单的体外伤口测定，以评估分层人角膜上皮细胞的愈合和屏障功能的恢复。在该测定中，圆形冲孔损伤导致上皮集体迁移为连贯片。伤口的闭合与跨细胞屏障的恢复和顶端细胞间连接的重建相关。总而言之，这种新的伤口愈合模型为研究导致复层上皮屏障功能的机制提供了重要的研究工具，并可能促进未来治疗应用的发展。
The repair of wounds through collective movement of epithelial cells is a fundamental process in multicellular organisms. In stratified epithelia such as the cornea and skin, healing occurs in three steps that include a latent, migratory, and reconstruction phases. Several simple and inexpensive assays have been developed to study the biology of cell migration in vitro. However, these assays are mostly based on monolayer systems that fail to reproduce the differentiation processes associated to multilayered systems. Here, we describe a straightforward in vitro wound assay to evaluate the healing and restoration of barrier function in stratified human corneal epithelial cells. In this assay, circular punch injuries lead to the collective migration of the epithelium as coherent sheets. The closure of the wound was associated with the restoration of the transcellular barrier and the re-establishment of apical intercellular junctions. Altogether, this new model of wound healing provides an important research tool to study the mechanisms leading to barrier function in stratified epithelia and may facilitate the development of future therapeutic applications.