title: Microfluidic device for DNA amplification of single cancer cells isolated from whole blood by self-seeding microwells.
abstract: Self-seeding microwell chips can sort single cells into 6400 wells based on cell size and their identity verified by immunofluorescence staining. Here, we developed a microfluidic device in which these single cells can be placed, lysed and their DNA amplified for further interrogation. Whole blood spiked with MCF7 tumor cells was passed through the microwell chips after leukocyte depletion and 37% of the MCF7 cells were identified by epithelial cell adhesion molecule (EpCAM) staining in the microwells. Identified single cells were punched into the reaction chamber of the microfluidic device and reagents for cell lysis and DNA amplification introduced sequentially by peristaltic pumping of micro-valves. On-chip lysis and amplification was performed in 8 parallel chambers yielding a 10,000 fold amplification of DNA. Accessibility of the sample through the reaction chamber allowed for easy retrieval and interrogation of target-specific genes to characterize the tumor cells.
authors.affiliation: Medical Cell BioPhysics Group, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, The Netherlands. l.w.m.m.terstappen@utwente.nl.
authors.forename: Yoonsun
authors.initials: Y
authors.lastname: Yang
authors.affiliation: Mesoscale Chemical Systems Group, MESA+ Institute for Nanotechnology, University of Twente, The Netherlands.
authors.forename: Hoon Suk
authors.initials: HS
authors.lastname: Rho
authors.affiliation: VyCAP, B.V., Deventer, The Netherlands.
authors.forename: Michiel
authors.initials: M
authors.lastname: Stevens
authors.affiliation: VyCAP, B.V., Deventer, The Netherlands.
authors.forename: Arjan G J
authors.initials: AG
authors.lastname: Tibbe
authors.affiliation: Mesoscale Chemical Systems Group, MESA+ Institute for Nanotechnology, University of Twente, The Netherlands.
authors.forename: Han
authors.initials: H
authors.lastname: Gardeniers
authors.affiliation: Medical Cell BioPhysics Group, MIRA Institute for Biomedical Technology and Technical Medicine, University of Twente, The Netherlands. l.w.m.m.terstappen@utwente.nl.
authors.forename: Leon W M M
authors.initials: LW
authors.lastname: Terstappen
mesh_terms: D000951:Antigens, Neoplasm / Q000032:analysis; D015815:Cell Adhesion Molecules / Q000032:analysis; D004273:DNA, Neoplasm / Q000235:genetics*; D000071858:Epithelial Cell Adhesion Molecule; D006801:Humans; D061986:MCF-7 Cells; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation; D059010:Single-Cell Analysis / Q000295:instrumentation
chemical_list: D000951:Antigens, Neoplasm; D015815:Cell Adhesion Molecules; D004273:DNA, Neoplasm; D000071858:Epithelial Cell Adhesion Molecule