High-throughput cell cycle synchronization using inertial forces in spiral microchannels |
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Authors: | Lee Wong Cheng Bhagat Ali Asgar S Huang Sha Van Vliet Krystyn J Han Jongyoon Lim Chwee Teck |
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Institution: | BioSystems and Micromechanics IRG, Singapore-MIT Alliance for Research and Technology Centre, Singapore. |
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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. |
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