Institut Fuer Analytische Chemie, Chemo- & Biosensorik, Universitaet Regensburg, Universitaetsstr. 31, 93053, Regensburg, Germany.
Ibidi GmbH, Lochhamer Schlag 11, 82166, Graefelfing, Germany.
Sci Rep. 2024 Apr 20;14(1):9103. doi: 10.1038/s41598-024-59564-9.
Quantitative assessment of cell migration in vitro is often required in fundamental and applied research from different biomedical areas including wound repair, tumor metastasis or developmental biology. A collection of assays has been established throughout the years like the most widely used scratch assay or the so-called barrier assay. It is the principle of these assays to introduce a lesion into an otherwise confluent monolayer in order to study the migration of cells from the periphery into this artificial wound and determine the migration rate from the time necessary for wound closure. A novel assay makes use of photosensitizers doped into a polystyrene matrix. A thin layer of this composite material is coated on the bottom of regular cell culture ware showing perfect biocompatibility. When adherent cells are grown on this coating, resonant excitation of the photosensitizer induces a very local generation of O, which kills the cells residing at the site of illumination. Cells outside the site of illumination are not harmed. When excitation of the photosensitizer is conducted by microscopic illumination, high-precision wounding in any size and geometry is available even in microfluidic channels. Besides proof-of-concept experiments, this study gives further insight into the mechanism of photosensitizer-mediated cell wounding.
体外细胞迁移的定量评估在不同的生物医学领域的基础和应用研究中经常需要，包括伤口修复、肿瘤转移或发育生物学。多年来已经建立了一系列的测定方法，如最广泛使用的划痕测定法或所谓的屏障测定法。这些测定法的原理是在原本致密的单层中引入一个损伤，以研究细胞从周围迁移到这个人工伤口，并从伤口闭合所需的时间来确定迁移率。一种新的测定法利用了掺杂在聚苯乙烯基质中的光增敏剂。这种复合材料的薄层涂覆在常规细胞培养器皿的底部，显示出极好的生物相容性。当附着的细胞在这种涂层上生长时，光增敏剂的共振激发会在非常局部的位置产生 O，杀死位于照明位置的细胞。而位于照明位置之外的细胞则不会受到伤害。当通过显微镜照明来激发光增敏剂时，即使在微流道中，也可以获得任何尺寸和形状的高精度损伤。除了概念验证实验，本研究还进一步深入了解了光增敏剂介导的细胞损伤的机制。
