Abstract 176: Generation and maintenance of anaplastic thyroid cancer spheroids from human cancer cells

Abstract Background: Anaplastic thyroid cancer (ATC) is the most aggressive thyroid cancer. Despite advances in tissue culture techniques, a robust model for thyroid cancer spheroid culture is yet to be developed. Here, we described a protocol to generate tumor spheroids from human ATC cells and excising cell lines. Methods: Using four established ATC cell lines (8505C, SW1736, C643 and THJ-16T) and one clinical tumor sample derived ATC cells, we created an efficient and cost-effective 3D culture system that can enhance our understanding of ex vivo treatment response. Results: We found that all four cell lines can readily form spheroids in culture with unique morphology, size, and cytoskeletal organization. We observed both cohesive (dense and solid structures) and dis-cohesive (irregularly shaped structures) spheroids within the same culture condition across different cell lines. BRAFWT ATC spheroids grew in a cohesive pattern, while BRAFV600E- mutant ATC spheroids show dis-cohesive organization. In the patient-derived spheroids (BRAFV600E- mutant ATC), we observed both growth patterns with mostly the dis-cohesive type. Phase-contrast images taken over time suggest that the cohesive ATC spheroids seem to be clonally derived, allowing the study of tumor heterogeneity. Furthermore, the ATC spheroids we established can maintain the 3D structures in the culture medium. Conclusion: Our study describes the development of a robust spheroid system from established ATC cell lines and freshly acquired patient tumor samples. We show that combining 3D culture with traditional 2D methods provides a complementary and powerful approach to the future drug screening and mechanism studies in the ATC. Citation Format: Jiangnan Hu, Chandrayee Ghosh, Electron Kebebew. Generation and maintenance of anaplastic thyroid cancer spheroids from human cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 176.