title: A digital microfluidic platform for primary cell culture and analysis.
abstract: Digital microfluidics (DMF) is a technology that facilitates electrostatic manipulation of discrete nano- and micro-litre droplets across an array of electrodes, which provides the advantages of single sample addressability, automation, and parallelization. There has been considerable interest in recent years in using DMF for cell culture and analysis, but previous studies have used immortalized cell lines. We report here the first digital microfluidic method for primary cell culture and analysis. A new mode of "upside-down" cell culture was implemented by patterning the top plate of a device using a fluorocarbon liftoff technique. This method was useful for culturing three different primary cell types for up to one week, as well as implementing a fixation, permeabilization, and staining procedure for F-actin and nuclei. A multistep assay for monocyte adhesion to endothelial cells (ECs) was performed to evaluate functionality in DMF-cultured primary cells and to demonstrate co-culture using a DMF platform. Monocytes were observed to adhere in significantly greater numbers to ECs exposed to tumor necrosis factor (TNF)-α than those that were not, confirming that ECs cultured in this format maintain in vivo-like properties. The ability to manipulate, maintain, and assay primary cells demonstrates a useful application for DMF in studies involving precious samples of cells from small animals or human patients.
authors.affiliation: Department of Mechanical & Industrial Engineering, University of Toronto, 5 King's College Road, Toronto, Ontario, Canada M5S 3G8.
authors.forename: Suthan
authors.initials: S
authors.lastname: Srigunapalan
authors.forename: Irwin A
authors.initials: IA
authors.lastname: Eydelnant
authors.forename: Craig A
authors.initials: CA
authors.lastname: Simmons
authors.forename: Aaron R
authors.initials: AR
authors.lastname: Wheeler
mesh_terms: D000818:Animals; D002448:Cell Adhesion; D002478:Cells, Cultured; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008856:Microscopy, Fluorescence; D009000:Monocytes / Q000166:cytology; D061251:Primary Cell Culture / Q000295:instrumentation*; D013552:Swine