From every glioma resection performed at Erasmus MC department of Neurosurgery, which provides sufficient tumor material, tissue is transported to the lab under an IRB-approved protocol. Part of the tissue is stored directly at -80 C for molecular analysis of the original patient material. Remaining tumor is dissociated (mechanically and enzymatically) using an optimized protocol* and is placed in defined serum-free culture medium*. The genotypic profile of the original tumor is preserved for at least 15 passages in derived cell cultures under these culture conditions.
*based on stringent analysis and comparison of multiple dissociation protocols and culture medium formulas.
Initially, tumor neurospheres are formed under serum-free culture conditions. These spheres are then transferred to wells containing various extracellular matrix coatings. The tumor cells attach to the most optimal substrate and grow in monolayer, allowing reproducible growth and viability assessment.
