title: Vacuum-assisted cell loading enables shear-free mammalian microfluidic culture.
abstract: Microfluidic perfusion cultures for mammalian cells provide a novel means for probing single-cell behavior but require the management of culture parameters such as flow-induced shear stress. Methods to eliminate shear stress generally focus on capturing cells in regions with high resistance to fluid flow. Here, we present a novel trapping design to easily and reliably load a high density of cells into culture chambers that are extremely isolated from potentially damaging flow effects. We utilize a transient on-chip vacuum to remove air from the culture chambers and rapidly replace the volume with a liquid cell suspension. We demonstrate the ability of this simple and robust method to load and culture three commonly used cell lines. We show how the incorporation of an on-chip function generator can be used for dynamic stimulation of cells during long-term continuous perfusion culture.
authors.affiliation: Department of Bioengineering, University of California, San Diego, CA, USA.
authors.forename: Martin
authors.initials: M
authors.lastname: Kolnik
authors.forename: Lev S
authors.initials: LS
authors.lastname: Tsimring
authors.forename: Jeff
authors.initials: J
authors.lastname: Hasty
mesh_terms: D000818:Animals; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000295:instrumentation*; D049109:Cell Proliferation; D006224:Cricetinae; D003412:Cricetulus; D005456:Fluorescent Dyes / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D055595:Mechanical Phenomena; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D014618:Vacuum
chemical_list: D005456:Fluorescent Dyes