To test the integrated system, we used a series of 3D tumor models developed at the Center for Radiological Research [53 (link)–55 (link)]. Human U87 Glioblastoma cells were chosen as an aggressive tumor model type known to grow rapidly in the 3D model system. These cells are representative of a hard-to-treat tumor that is a prime candidate for HIRT. As described previously [56 (link)], to form 3D cell cultures, human U87 Glioblastoma cells were grown as a monolayer, trypsinized, injected into the gel matrix and incubated for 24-48 h before imaging. Depending on the cell type, these injected cell boluses can form either a solid tumor core or a more dispersed cell distribution. U87’s high motility and potential for tissue invasion form a more dispersed cell bolus.
We tested the combined imaging and irradiation system with a range of cell labelling strategies: Cyto-Red (Biosettia) and CellTracker Red (Thermo Fisher) for whole cell labeling, and Oregon Green BAPTA-1 AM (Thermo Fisher), a calcium sensitive fluorescent dye as a functional reporter cellular Ca2+ dynamics. We also tested human U87 cells transfected with the pGreenFire 2.0 NFkB reporter (SBI).
We tested the combined imaging and irradiation system with a range of cell labelling strategies: Cyto-Red (Biosettia) and CellTracker Red (Thermo Fisher) for whole cell labeling, and Oregon Green BAPTA-1 AM (Thermo Fisher), a calcium sensitive fluorescent dye as a functional reporter cellular Ca2+ dynamics. We also tested human U87 cells transfected with the pGreenFire 2.0 NFkB reporter (SBI).
