Yu Ling-Yea, Lin Hsuan-Chao, Hsu Chi-Lin, Kao Tuan-Yu, Tsai Feng-Chiao
Department of Pharmacology, National Taiwan University College of Medicine, Taipei, Taiwan.
Department of Internal Medicine, National Taiwan University Hospital, Taipei, Taiwan.
Bio Protoc. 2023 Nov 5;13(21):e4871. doi: 10.21769/BioProtoc.4871.
Cell migration is an essential biological process for organisms, in processes including embryonic development, immune response, and cancer metastasis. To elucidate the regulatory machinery of this vital process, methods that mimic in vivo migration, including in vitro wound healing assay and random migration assay, are widely used for cell behavior investigation. However, several concerns are raised with traditional cell migration experiment analysis. First, a manually scratched wound often presents irregular edges, causing the speed analysis difficult. Second, only the migration speed of leading cells is considered in the wound healing assay. Here, we provide a reliable analysis method to trace each cell in the time-lapse images, eliminating the concern about wound shape and creating a more comprehensive understanding of cell migration-not only of collective migration speed but also single-cell directionality and coordination between cells.
细胞迁移是生物体的一个基本生物学过程，存在于包括胚胎发育、免疫反应和癌症转移等过程中。为了阐明这一重要过程的调控机制，模拟体内迁移的方法，包括体外伤口愈合试验和随机迁移试验，被广泛用于细胞行为研究。然而，传统的细胞迁移实验分析引发了一些问题。首先，人工划痕造成的伤口边缘往往不规则，导致速度分析困难。其次，伤口愈合试验中仅考虑了前沿细胞的迁移速度。在此，我们提供了一种可靠的分析方法，用于在延时图像中追踪每个细胞，消除了对伤口形状的担忧，并对细胞迁移有了更全面的理解——不仅包括集体迁移速度，还包括单细胞方向性和细胞间的协调性。
