title: A Microfluidic Platform for High-throughput Single-cell Isolation and Culture.
abstract: 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 (~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.
authors.affiliation: Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Taiwan; Tissue Engineering and Regenerative Medicine, National Chung Hsing University.
authors.forename: Ching-Hui
authors.initials: CH
authors.lastname: Lin
authors.affiliation: Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Taiwan; Tissue Engineering and Regenerative Medicine, National Chung Hsing University.
authors.forename: Hao-Chen
authors.initials: HC
authors.lastname: Chang
authors.affiliation: Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Taiwan; Tissue Engineering and Regenerative Medicine, National Chung Hsing University; Institute of NanoEngineering and MicroSystems, National Tsing Hua University; chsu@nhri.org.tw.
authors.forename: Chia-Hsien
authors.initials: CH
authors.lastname: Hsu
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002469:Cell Separation; D006801:Humans; D051379:Mice; D046210:Microfluidic Analytical Techniques; D044085:Microfluidics; D058953:Neural Stem Cells