title: A microfluidic dual-well device for high-throughput single-cell capture and culture.
abstract: 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.
authors.affiliation: Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, 35 Keyan Road, Zhunan, Miaoli County 35053, Taiwan. chsu@nhri.org.tw.
authors.forename: Ching-Hui
authors.initials: CH
authors.lastname: Lin
authors.forename: Yi-Hsing
authors.initials: YH
authors.lastname: Hsiao
authors.forename: Hao-Chen
authors.initials: HC
authors.lastname: Chang
authors.forename: Chuan-Feng
authors.initials: CF
authors.lastname: Yeh
authors.forename: Cheng-Kun
authors.initials: CK
authors.lastname: He
authors.forename: Eric M
authors.initials: EM
authors.lastname: Salm
authors.forename: Chihchen
authors.initials: C
authors.lastname: Chen
authors.forename: Ing-Ming
authors.initials: IM
authors.lastname: Chiu
authors.forename: Chia-Hsien
authors.initials: CH
authors.lastname: Hsu
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D057166:High-Throughput Screening Assays / Q000295:instrumentation; D006801:Humans; D051379:Mice; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D058953:Neural Stem Cells / Q000166:cytology*; D059010:Single-Cell Analysis / Q000295:instrumentation