title: High-throughput cell cycle synchronization using inertial forces in spiral microchannels.
abstract: Efficient synchronization and selection of cells at different stages of the cell replication cycle facilitates both fundamental research and development of cell cycle-targeted therapies. Current chemical-based synchronization methods are unfavorable as these can disrupt cell physiology and metabolism. Microfluidic systems developed for physical cell separation offer a potential alternative over conventional cell synchronization approaches. Here we introduce a spiral microfluidic device for cell cycle synchronization, using the combined effects of inertial forces and Dean drag force. By exploiting the relationship between cell diameter and cell cycle (DNA content/ploidy), we have successfully fractionated several asynchronous mammalian cell lines, as well as primary cells comprising bone marrow-derived human mesenchymal stem cells (hMSCs), into enriched subpopulations of G0/G1 (>85%), S, and G2/M phases. This level of cell cycle enrichment is comparable to existing microfluidic systems, but the throughput (∼ 15 × 10(6) cells per h) and viability (∼ 95%) of cells thus synchronized are significantly greater. Further, this platform provides rapid collection of synchronized cells or of diameter-sorted cells post-separation, to enable diverse applications in the study and manipulation of cell proliferation.
authors.affiliation: BioSystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology Centre, Singapore.
authors.forename: Wong Cheng
authors.initials: WC
authors.lastname: Lee
authors.forename: Ali Asgar S
authors.initials: AA
authors.lastname: Bhagat
authors.forename: Sha
authors.initials: S
authors.lastname: Huang
authors.forename: Krystyn J
authors.initials: KJ
authors.lastname: Van Vliet
authors.forename: Jongyoon
authors.initials: J
authors.lastname: Han
authors.forename: Chwee Teck
authors.initials: CT
authors.lastname: Lim
mesh_terms: D000818:Animals; D016466:CHO Cells; D002453:Cell Cycle; D002469:Cell Separation / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D006367:HeLa Cells; D006801:Humans; D055595:Mechanical Phenomena; D059630:Mesenchymal Stem Cells / Q000166:cytology; D046210:Microfluidic Analytical Techniques / Q000379:methods*