Laboratoire Physico-chimie Curie, Institut Curie, Centre de Recherche, CNRS, Université Pierre et Marie Curie, Paris, France.
Biophys J. 2010 May 19;98(9):1790-800. doi: 10.1016/j.bpj.2010.01.030.
We report quantitative measurements of the velocity field of collectively migrating cells in a motile epithelium. The migration is triggered by presenting free surface to an initially confluent monolayer by using a microstencil technique that does not damage the cells. To avoid the technical difficulties inherent in the tracking of single cells, the field is mapped using the technique of particle image velocimetry. The main relevant parameters, such as the velocity module, the order parameter, and the velocity correlation function, are then extracted from this cartography. These quantities are dynamically measured on two types of cells (collectively migrating Madin-Darby canine kidney (MDCK) cells and fibroblastlike normal rat kidney (NRK) cells), first as they approach confluence, and then when the geometrical constraints are released. In particular, for MDCK cells filling up the patterns, we observe a sharp decrease in the average velocity after the point of confluence, whereas the densification of the monolayer is much more regular. After the peeling off of the stencil, a velocity correlation length of approximately 200 microm is measured for MDCK cells versus only approximately 40 microm for the more independent NRK cells. Our conclusions are supported by parallel single-cell tracking experiments. By using the biorthogonal decomposition of the velocity field, we conclude that the velocity field of MDCK cells is very coherent in contrast with the NRK cells. The displacements in the fingers arising from the border of MDCK epithelia are very oriented along their main direction. They influence the velocity field in the epithelium over a distance of approximately 200 microm.
我们报告了在一个运动上皮中集体迁移细胞的速度场的定量测量。迁移是通过使用微掩模技术将自由表面呈现给最初汇合的单层来触发的,该技术不会损坏细胞。为了避免跟踪单个细胞所固有的技术困难,使用粒子图像测速技术来绘制该场。然后,从该映射中提取主要相关参数,例如速度模块、序参量和速度相关函数。这些数量在两种类型的细胞(集体迁移的 Madin-Darby 犬肾 (MDCK) 细胞和类成纤维细胞的正常大鼠肾 (NRK) 细胞)上进行动态测量,首先是在它们接近汇合时,然后是在释放几何约束时。特别是,对于填充图案的 MDCK 细胞,我们观察到在汇合点之后平均速度急剧下降,而单层的密集化则更加规则。掩模剥落后,对于 MDCK 细胞测量到大约 200 µm 的速度相关长度,而对于更加独立的 NRK 细胞仅测量到大约 40 µm。我们的结论得到了平行的单细胞跟踪实验的支持。通过使用速度场的双正交分解,我们得出结论,与 NRK 细胞相比,MDCK 细胞的速度场非常一致。源自 MDCK 上皮边缘的手指的位移非常沿着它们的主要方向定向。它们在大约 200 µm 的距离内影响上皮中的速度场。