High resolution time‐lapse analysis of glioma tumor cell migration

Abstract

Methods generally in use to measure tumor cell movements lack sufficiency in numbers of cells analyzed, precision of measurements, or temporal and spatial resolution. Here, the study of glioma cell motility was greatly enhanced using a fully automated time-lapse microscopy system capable of collecting and analyzing motility data at closely spaced time points (5 min), over long periods (24 hrs), and under several different experimental conditions in parallel. This system was designed to be significantly more versatile and less costly than commercial systems and collected data under phase contrast and widefield fluorescent illumination concurrently. Human and rat glioma cell lines were plated under a variety of conditions and subjected to time-lapse microscopy, cell tracking, and quantitative analysis of velocity, total accumulated distance, and directionality for individual cells or for averaged cell populations. Quantitation of glioma “scratch” assays revealed changes in motility parameters after 1) anti-adhesion molecule antibody-treatment, 2) adhesion molecule-transfection, or 3) antisense-adhesion molecule viral vector infection. Fluorescently labeled glioma cells were tracked while migrating on top of cell monolayers that expressed ectopic adhesion molecules, and this resulted in significantly reduced glioma migration velocities. Our methods of analysis revealed changes in glioma cell motility after experimental treatments that would not be discernable by other common methods. Supported by the HHMI Undergraduate Research Program, the McNair Scholars Program, and the U.D. Undergraduate Research Program.