title: CycleTrak: a novel system for the semi-automated analysis of cell cycle dynamics.
abstract: Cell proliferation is crucial to tissue growth and form during embryogenesis, yet dynamic tracking of cell cycle progression and cell position presents a challenging roadblock. We have developed a fluorescent cell cycle indicator and single cell analysis method, called CycleTrak, which allows for better spatiotemporal resolution and quantification of cell cycle phase and cell position than current methods. Our method was developed on the basis of the existing Fucci method. CycleTrak uses a single lentiviral vector that integrates mKO2-hCdt1 (30/120), and a nuclear-localized eGFP reporter. The single vector and nuclear localized fluorescence signals simplify delivery into cells and allow for rapid, automated cell tracking and cell cycle phase readout in single and subpopulations of cells. We validated CycleTrak performance in metastatic melanoma cells and identified novel cell cycle dynamics in vitro and in vivo after transplantation and 3D confocal time-lapse imaging in a living chick embryo.
authors.affiliation: Stowers Institute for Medical Research, 1000 E. 50th St., Kansas City, MO 64110, USA.
authors.forename: Dennis A
authors.initials: DA
authors.lastname: Ridenour
authors.forename: Mary Cathleen
authors.initials: MC
authors.lastname: McKinney
authors.forename: Caleb M
authors.initials: CM
authors.lastname: Bailey
authors.forename: Paul M
authors.initials: PM
authors.lastname: Kulesa
mesh_terms: D000818:Animals; D002453:Cell Cycle; D058948:Cell Tracking / Q000379:methods*; D002642:Chick Embryo; D005453:Fluorescence; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins; D006367:HeLa Cells; D006801:Humans; D016086:Lentivirus; D059010:Single-Cell Analysis / Q000379:methods*; D059008:Time-Lapse Imaging
chemical_list: D049452:Green Fluorescent Proteins