TMOD-14. INNOVATIVE 3D MODEL FOR THE ESTABLISHMENT OF PRIMARY PAEDIATRIC LOW-GRADE GLIOMA (LGG) CULTURES: NEW PLATFORM FOR ADVANCED PRECLINICAL STUDIES OF INNOVATIVE AND IMMUNOTHERAPEUTIC APPROACHES

• Published in: Neuro-oncology (Charlottesville, Va.) Va.), 2019-04, Vol.21 (Supplement_2), p.ii123-ii124
• Main Authors: Belardinilli, Tamascia, Arnone, Claudia Manuela, Mastronuzzi, Angela, Ceglie, Giulia, Cacchione, Antonella, Carai, Andrea, Miele, Evelina, Camassei, Francesca Diomedi, Polito, Vinicia Assunta, Stefanis, Cristiano De, Quintarelli, Concetta, Angelis, Biagio De, Locatelli, Franco, Caruana, Ignazio, Del Bufalo, Francesca
• Format: Article
• Language: English
• Subjects: Pediatric Brain Tumor Models
• ISSN: 1522-8517; 1523-5866
• DOI: 10.1093/neuonc/noz036.251

Abstract Solid tumours are characterized by a high structural complexity that is difficult to reproduce with common bidimensional (2D) systems. We developed an innovative fibrin-based hydrogel 3D model and tested it as platform to establish primary LGG cultures, otherwise difficult to maintain owing to the activation of senescence pathways. To date, 37 samples were cultured in both 2D and 3D platforms (19 astrocytomas and 18 gangliogliomas), with an average culture duration of 177+13 days, showing that the 3D-culture enables the stabilization of LGG. Whereas in the 2D-culture the cells showed overtime a significant reduction in the average culture days between each passage (average P1-P3:35+6 days; P3-P6: 29+5; >P6: 17+5), suggesting the selection of a peculiar clone, in the 3D-setting, we observed a more regular growth (average P1-P3:44+5 days; P3-P6: 64+20;>P6: 48+16). Cell lines identity was confirmed by short tandem repeats (STRs) and the immunohistochemical characterization (H&E, Ki67, tumour and differentiation markers) in 3D cultures revealed phenotype, cellular organization and proliferative rates closer to those observed in the onset samples, as compared to the 2D. We then analyzed the methylation profile and preliminary results suggest that the 2D lines evolve towards different lineages. The analysis of cell senescence using ß-galactosidase assay revealed a lower senescence in the 3D cultures (3D:12,28%+4,3% of the cells vs 2D:50,69%+18,46%; p=0,008). Lastly, we evaluated the responses to radiotherapy by MTS assay, and showed that 2D-cultured cells are more sensitive to treatment than 3D (2D: untreated, 0,0232+0,023, 160 Gy 0,089+0,025; 3D: untreated 0,235+0,09, 160 Gy 0,169+0,08), suggesting an overestimation of the efficacy of such treatment by the 2D-setting, therefore reducing the predictive power. The response to chemotherapy and more innovative approaches (i.e. Oncolytic Adenovirus) is currently under evaluation, the preliminary results confirming a similar finding. Overall, the 3D-culture offers an innovative platform for biological and therapeutic studies.