13C NMR Study of Caffeine Metabolism in Glioblastoma and NHAs with Chemotherapy*

The use of 3D spheroids in conjunction with traditional 2D cell culture methods is essential for pre-clinical studies aimed at understanding glioblastoma development. Spheroids more accurately replicate tumor architecture, cell-to-cell interactions, and the tumor microenvironment compared to conventional 2D cultures, making them a superior model for cancer research. In this study, we compared 2D monolayer and 3D spheroid cultures of glioblastoma U87 and normal human astrocyte (NHA) cell lines, utilizing ultra-low attachment (ULA) plates and the hanging drop method. Previous studies have shown that caffeine inhibits monolayer U87 growth, sensitizing glioblastoma cells to chemotherapy, specifically temozolomide. This research seeks to determine whether this effect extends to U87 spheroids when treated with carboplatin and caffeine. The results of this project will be discussed considering subsequent 13C NMR spectroscopic studies to probe aberrant metabolism in cancer cells using the spheroid models. Further research has scope to test this combination’s effect on healthy brain cells and normal human astrocytes.