摘要
Single-cell culture is a method of growing isolated single-cell routinely performed to obtain single-cell-derived cell clones for both basic research and therapeutically applications. Besides forming clones, culturing individually isolated single cells can also help understand the metabolic, migration and differentiation heterogeneity in cell populations. Despite single-cell culture being essential and widely used, single-cell culture is still a challenging task for many laboratories due to the difficulty of manipulating and observing single cells. Microfluidic techniques have emerged as a useful tool for single-cell culture applications. In comparison to conventional petri dishes and well plates, culturing cells in microfluidic devices offers several advantages because: (1) the microdevices can more precisely manipulate single cells and control the cell culture conditions, useful for providing better controlled experimental conditions. For example, microfluidic channel can be used to generate stable chemical gradients for studying single-cell chemotaxis at high temporal and/or spatial resolution. (2) The miniaturized devices can increase the throughput and reduce reagent/cell consumptions of experiments due to their small sizes. And (3) microfluidic devices are amenable to integration with microelectromechanical systems (MEMS) technology to form lab-on-a-chip devices which allow for combining single-cell culture and analysis in one device. Besides making cell-culture devices, microfluidic techniques can also be used as a helper tool to prepare cells for single-cell culture with conventional culture dishes and well plates for applications where single-cell suspension preparation is difficult. In this chapter, state-of-the-art microfluidic techniques for single-cell preparation, separation and culture will be described and compared.