A Simple 3D Cell Culture Method for Studying the Interactions between Human Mesenchymal Stromal/Stem Cells and Patients Derived Glioblastoma †

SIMPLE SUMMARY: Glioblastoma (GBM) remain an incurable disease despite important efforts to find efficient treatments. GBM are heterogeneous, complex, and plastic tumors and as such it is expected that personalized treatments should be the key to therapeutic success. It is thus of the utmost importance to design new methods to mirror the GBM progression and response to treatments for each patient. Our results show that the co-culture of primary cultures derived from GBM patients with cancer associated fibroblasts derived from mesenchymal stem cells exhibit in vitro growth and morphology close to that found in vivo with patients. ABSTRACT: We have developed a 3D biosphere model using patient-derived cells (PDCs) from glioblastoma (GBM), the major form of primary brain tumors in adult, plus cancer-activated fibroblasts (CAFs), obtained by culturing mesenchymal stem cells with GBM conditioned media. The effect of MSC/CAFs on the proliferation, cell-cell interactions, and response to treatment of PDCs was evaluated. Proliferation in the presence of CAFs was statistically lower but the spheroids formed within the 3D-biosphere were larger. A treatment for 5 days with Temozolomide (TMZ) and irradiation, the standard therapy for GBM, had a marked effect on cell number in monocultures compared to co-cultures and influenced cancer stem cells composition, similar to that observed in GBM patients. Mathematical analyses of spheroids growth and morphology confirm the similarity with GBM patients. We, thus, provide a simple and reproducible method to obtain 3D cultures from patient-derived biopsies and co-cultures with MSC with a near 100% success. This method provides the basis for relevant in vitro functional models for a better comprehension of the role of tumor microenvironment and, for precision and/or personalized medicine, potentially to predict the response to treatments for each GBM patient.