Researcher(s)
- Zachary Waterman, Biological Sciences, University of Delaware
Faculty Mentor(s)
- Deni Galileo, Biology, University of Delaware
Abstract
Glioblastoma (GBM) has the highest invasion and growth rates of all brain cancers in adults and is classified as a grade IV malignant tumor by the WHO. Previously, the Galileo lab performed xenograft experiments in chick embryo brains to study invasiveness of GBM cell lines and the role of the L1CAM adhesion protein. Over the past year, procedures were established for introducing small GBM cell line aggregates onto the chick embryo brain slices (i.e., ex vivo model). Additionally, a previously published senior thesis student paper established the effectiveness of certain inhibitor drug treatments on GBM cell motility and proliferation in a dish, including Y15, Cilengitide, PD173074, and PF431396. My summer experiments focused on combining these two procedures to determine the effectiveness of the inhibitors on GBM cell aggregates in brain slices. To do this, embryonic day (E) 15 brains were dissected and sectioned into 350-micron slices. Brains slices were cultured in appropriate media for three or six days, at which point GBM cell aggregates were introduced onto the slices. Red fluorescent U-118/mCherry GBM cells that either expressed the L1CAM adhesion protein ectodomain or did not, were introduced onto the brain slice as aggregates. After allowing a day for aggregates to attach, the slices were treated with one of the aforementioned drugs for a period of one to two days. Brain slices were then fixed and immunostained for laminin surrounding blood vessels and counterstained for nuclei. Red GBM cell invasion was assessed using the grid-based analysis in which cells in a grid are scored either positive or negative for glioblastoma cells. Analysis of GBM cell invasion in the brain slices is ongoing to determine the best time frame for treatment and if significant differences exist between drug treatments and controls. I expect the three-day attachment period and subsequent two-day drug treatment will lead to significant differences between drug treatments and control conditions.
Supported by a grant from the Lisa Dean Moseley Foundation.