Recovery of glioblastoma cells derived from patients after a decade of cryopreservation for drug response studies.

Patient-derived glioblastoma cells (GBMs) are essential for drug screening and personalized medicine. Limited patient sample availability requires the use of long-term cryopreserved GBM cells. However, current methods for recovering GBM cells after an extended period of cryopreservation are generally inefficient, resulting in poor survival of these cells. Therefore, this study sought to optimize a culture procedure to improve the recovery and growth of patient-derived GBM cells after long-term cryostorage. We show that the use of Matrigel with an increased percentage of fetal bovine serum significantly improves the viability and subsequent expansion of GBM cells, which were cryopreserved for more than 10 years, compared to standard culture methods. The enhanced growth is associated with an increase in the expression of YAP and TLR4, the key regulators of cell proliferation, in these GBM cells. These recovered GBM cells were successfully used for chemotherapeutic drug testing, specifically temozolomide (TMZ), in 2D and 3D spheroid culture systems. We believe that our optimized protocol offers a valuable tool to increase the availability of patient-derived GBM cells by allowing efficient recovery of long-term cryopreserved GBM cells for research and personalized drug testing.