A Microfluidic Platform for High-throughput Single-cell Isolation and Culture

作者：Lin Ching Hui; Chang Hao Chen; Hsu Chia Hsien

来源：Jove-Journal of Visualized Experiments, 2016, (112): e54105.

DOI：10.3791/54105

• Bioengineering
• Issue 112
• Microfluidics
• High-throughput
• High-efficiency
• Single cell
• Cell isolation
• Cell culture
• Clonogenic assay

Studying the heterogeneity of single cells is crucial for many biological questions, but is technically difficult. Thus, there is a need for a simple, yet high-throughput, method to perform single-cell culture experiments. Here, we report a microfluidic chip-based strategy for high-efficiency single-cell isolation (similar to 77%) and demonstrate its capability of performing long-term single-cell culture (up to 7 d) and cellular heterogeneity analysis using clonogenic assay. These applications were demonstrated with KT98 mouse neural stem cells, and A549 and MDA-MB-435 human cancer cells. High single-cell isolation efficiency and long-term culture capability are achieved by using different sizes of microwells on the top and bottom of the microfluidic channel. The small microwell array is designed for precisely isolating single-cells, and the large microwell array is used for single-cell clonal culture in the microfluidic chip. This microfluidic platform constitutes an attractive approach for single-cell culture applications, due to its flexibility of adjustable cell culture spaces for different culture strategies, without decreasing isolation efficiency.

• 出版日期2016-6
• 单位清华大学