Glioblastoma Multiforme (GBM) is a common and deadly brain tumor that affects 10,000-12,000 people per year. There is currently no cure for GBM, and standard cancer treatments such as surgical operations, radiation, and chemotherapy are ineffective against this disease. Studies have been conducted to help further the efficacy of treatments for GBM, however, the necessity to further understand GBM’s molecular biology has become quite evident. Autobioluminescence is the chemical self-production of light using an endogenously-supplied (self-made) substrate reacting with an enzyme and is widely used for molecular imaging purposes. Autobioluminescence is especially advantageous in the imaging field, as it gives the sensitivity and longitudinal imaging that is not provided by other imaging techniques, such as traditional bioluminescence and fluorescence. In order to advance the understanding of GBM’s molecular biology, we plan to develop autobioluminescent patient-derived GBM cellular models and then characterize the models’ applications for high-throughput screening assays (high-speed drug screening techniques) in 2D, 3D, and 3D astrocyte (sub-type of glial cell in the brain) co-culture assays. Firstly, we will create the cellular models using three patient-derived GBM cell lines which are HF2381, HF3013, and HF3177 Secondly, we will evaluate our GBM cellular models in 2D, 3D, and 3D astrocyte spheroid (environment is in the shape of a sphere for better visualization) co-culture environment. Lastly, we will create an autobioluminescent GBM cell and endothelial cell (a type of cell found in blood vessels and the heart) co-culture model to visualize the invasion of GBM in vitro (outside of an organism). The results of this study, upon successful completion, will consist of three autobioluminescent patient-derived GBM cellular models and will have evaluated the models’ response profiles in 2D, 3D, and 3D astrocyte co-culture environments.