Donath Sören, Seidler Anna Elisabeth, Mundin Karlina, Wenzel Johannes, Scholz Jonas, Gentemann Lara, Kalies Julia, Faix Jan, Ngezahayo Anaclet, Bleich André, Heisterkamp Alexander, Buettner Manuela, Kalies Stefan
Institute of Quantum Optics, Leibniz University Hannover, 30167 Hannover, Germany.
Lower Saxony Center for Biomedical Engineering, Implant Research and Development (NIFE), 30625 Hannover, Germany.
iScience. 2023 Oct 5;26(11):108139. doi: 10.1016/j.isci.2023.108139. eCollection 2023 Nov 17.
Intestinal organoids represent a three-dimensional cell culture system mimicking the mammalian intestine. The application of single-cell ablation for defined wounding via a femtosecond laser system within the crypt base allowed us to study cell dynamics during epithelial restitution. Neighboring cells formed a contractile actin ring encircling the damaged cell, changed the cellular aspect ratio, and immediately closed the barrier. Using traction force microscopy, we observed major forces at the ablation site and additional forces on the crypt sides. Inhibitors of the actomyosin-based mobility of the cells led to the failure of restoring the barrier. Close to the ablation site, high-frequency calcium flickering and propagation of calcium waves occured that synchronized with the contraction of the epithelial layer. We observed an increased signal and nuclear translocation of YAP-1. In conclusion, our approach enabled, for the first time, to unveil the intricacies of epithelial restitution beyond models by employing precise laser-induced damage in colonoids.
肠道类器官代表了一种模拟哺乳动物肠道的三维细胞培养系统。通过飞秒激光系统在隐窝底部进行特定创伤的单细胞消融，使我们能够研究上皮修复过程中的细胞动态。相邻细胞形成围绕受损细胞的收缩性肌动蛋白环，改变细胞长宽比，并立即封闭屏障。使用牵引力显微镜，我们观察到消融部位的主要力以及隐窝侧面的附加力。基于肌动球蛋白的细胞迁移抑制剂导致屏障恢复失败。在消融部位附近，出现了高频钙闪烁和钙波传播，这与上皮层的收缩同步。我们观察到YAP-1的信号增加和核转位。总之，我们的方法首次通过在类结肠中进行精确的激光诱导损伤，揭示了上皮修复过程中超出模型的复杂性。
