Functional Interplay Between Cell Cycle and Cell Phenotypes

Cell cycle distribution of adherent cells is typically assessed using flow cytometry following detachment of the cells, which presumes the measurements of cell properties (cell size and nuclear size) and cell cycle distribution are still the same in the original environment. Here we use a high-throughput microscopy system to simultaneously analyze the cell-cycle phase of thousands of individual cells and their associated cell properties. This assay demonstrates that cell properties strongly depend on cell-cycle phase. By perturbing cell cycle with synchronization or inhibition of cell cycle regulator or changing nuclear size and shape with knockdown structural proteins (Nesprins, Lamin A/C), cell cycle and cell phenotypes always interfere with each other. With a linear combination equation, we could calculate how much effect on phenotypes is due to cell cycle redistribution or intrinsic phenotypes. This study highlights the functional interplay between cell cycle and cell phenotypes by showing the simultaneous, in situ measurements of cell cycle and phenotypes at single-cell resolution, a method that could be used in a wide range of applications in biology and biomedical research.