School of Mechanical Engineering, Korea University, Seoul, Korea.
Nat Protoc. 2012 Jun 7;7(7):1247-59. doi: 10.1038/nprot.2012.051.
This protocol describes a simple but robust microfluidic assay combining three-dimensional (3D) and two-dimensional (2D) cell culture. The microfluidic platform comprises hydrogel-incorporating chambers between surface-accessible microchannels. By using this platform, well-defined biochemical and biophysical stimuli can be applied to multiple cell types interacting over distances of <1 mm, thereby replicating many aspects of the in vivo microenvironment. Capabilities exist for time-dependent manipulation of flow and concentration gradients as well as high-resolution real-time imaging for observing spatial-temporal single-cell behavior, cell-cell communication, cell-matrix interactions and cell population dynamics. These heterotypic cell type assays can be used to study cell survival, proliferation, migration, morphogenesis and differentiation under controlled conditions. Applications include the study of previously unexplored cellular interactions, and they have already provided new insights into how biochemical and biophysical factors regulate interactions between populations of different cell types. It takes 3 d to fabricate the system and experiments can run for up to several weeks.
本方案描述了一种简单但强大的微流控分析方法，结合了三维（3D）和二维（2D）细胞培养。微流控平台包括在可接近表面的微通道之间的水凝胶结合室。通过使用该平台，可以对多种细胞类型施加明确定义的生化和生物物理刺激，这些细胞类型在<1mm 的距离上相互作用，从而复制体内微环境的许多方面。该平台具有时间依赖性的流动和浓度梯度的操控能力，以及用于观察时空单细胞行为、细胞-细胞通讯、细胞-基质相互作用和细胞群体动力学的高分辨率实时成像能力。这些异质细胞类型的分析可以用于研究细胞在受控条件下的存活、增殖、迁移、形态发生和分化。该方法的应用包括研究以前未被探索的细胞相互作用，并且已经为理解生化和生物物理因素如何调节不同细胞类型群体之间的相互作用提供了新的见解。该系统的制作需要 3 天时间，实验可以持续数周。
