Vargas Pablo, Terriac Emmanuel, Lennon-Duménil Ana-Maria, Piel Matthieu
Immunité et Cancer, Institut Curie.
Compartimentation et Dynamique Cellulaires, Institut Curie.
J Vis Exp. 2014 Feb 21(84):e51099. doi: 10.3791/51099.
The method described here allows the study of cell migration under confinement in one dimension. It is based on the use of microfabricated channels, which impose a polarized phenotype to cells by physical constraints. Once inside channels, cells have only two possibilities: move forward or backward. This simplified migration in which directionality is restricted facilitates the automatic tracking of cells and the extraction of quantitative parameters to describe cell movement. These parameters include cell velocity, changes in direction, and pauses during motion. Microchannels are also compatible with the use of fluorescent markers and are therefore suitable to study localization of intracellular organelles and structures during cell migration at high resolution. Finally, the surface of the channels can be functionalized with different substrates, allowing the control of the adhesive properties of the channels or the study of haptotaxis. In summary, the system here described is intended to analyze the migration of large cell numbers in conditions in which both the geometry and the biochemical nature of the environment are controlled, facilitating the normalization and reproducibility of independent experiments.
这里描述的方法允许在一维受限条件下研究细胞迁移。它基于使用微制造通道，这些通道通过物理约束赋予细胞极化表型。一旦进入通道，细胞只有两种可能性：向前移动或向后移动。这种方向性受限的简化迁移便于自动跟踪细胞并提取描述细胞运动的定量参数。这些参数包括细胞速度、方向变化和运动过程中的停顿。微通道也与荧光标记物的使用兼容，因此适合在高分辨率下研究细胞迁移过程中细胞内细胞器和结构的定位。最后，通道表面可以用不同的底物进行功能化，从而控制通道的粘附特性或研究趋触性。总之，这里描述的系统旨在分析在环境的几何形状和生化性质均受到控制的条件下大量细胞的迁移，有助于独立实验的标准化和可重复性。
