A microfluidic dual-well device for high-throughput single-cell capture and culture

• Published in: Lab on a chip Vol. 15; no. 14; pp. 2928 - 2938
• Main Authors: Lin, Ching-Hui, Hsiao, Yi-Hsing, Chang, Hao-Chen, Yeh, Chuan-Feng, He, Cheng-Kun, Salm, Eric M., Chen, Chihchen, Chiu, Ing-Ming, Hsu, Chia-Hsien
• Format: Journal Article
• Language: English
• Published: England 01.01.2015
• Subjects: Animals; Biotechnology; Cell Differentiation; Cell Proliferation; Cells, Cultured; Culture; Devices; Differentiation; Heterogeneity; High-Throughput Screening Assays - instrumentation; Humans; Mice; Microfluidic Analytical Techniques - instrumentation; Microfluidics; Neural Stem Cells - cytology; Single-Cell Analysis - instrumentation; Spreading; Stem cells
• ISSN: 1473-0197; 1473-0189
• DOI: 10.1039/C5LC00541H

In vitro culture of single cells facilitates biological studies by deconvoluting complications from cell population heterogeneity. However, there is still a lack of simple yet high-throughput methods to perform single cell culture experiments. In this paper, we report the development and application of a microfluidic device with a dual-well (DW) design concept for high-yield single-cell loading (~77%) in large microwells (285 and 485 μm in diameter) which allowed for cell spreading, proliferation and differentiation. The increased single-cell loading yield is achieved by using sets of small microwells termed “capture-wells” and big microwells termed “culture-wells” according to their utilities for single-cell capture and culture, respectively. This novel device architecture allows the size of the “culture” microwells to be flexibly adjusted without affecting the single-cell loading efficiency making it useful for cell culture applications as demonstrated by our experiments of KT98 mouse neural stem cell differentiation, A549 and MDA-MB-435 cancer cell proliferation, and single-cell colony formation assay with A549 cells in this paper.