Hyaluronic-Acid Based Hydrogels for 3-Dimensional Culture of Patient-Derived Glioblastoma Cells

This method can help answer key questions about glioblastoma tumors and how the extracellular matrix can facilitate treatment resistance. This technique enables modular construction of adaptable 3D cell culture platforms which can be used to culture patient-derived glioblastoma cells in an environment that better approximates native brain. To begin, place clean dry silicone rubber molds into each well of a non-tissue culture treated 12-well plate and use the clean blunt end of a pipette tip to adhere the molds to the bottom of the plate.

After checking the seal between the molds and the well bottoms, collect the dissociated gliomaspheres from a glioblastoma cell culture by centrifugation and re-suspend the pellet in one milliliter of cell dissociation enzyme. After five minutes at room temperature, agitate the tube with gentle tapping and arrest the enzymatic reaction with four milliliters of complete medium. Centrifuge the cells again and re-suspend the pellet in one milliliter of fresh complete medium before straining the cells through a 70 micrometer filter.