3D in vitro models of head & neck squamous cell carcinomas carcinomas using patient-derived cells

Introduction: Head and neck squamous cell carcinomas (HNSCCs) have a yearly incidence of 600,000 cases worldwide, with a mortality rate of 40%–50%. As a result, techniques to better understand the HNSCCs and, eventually, develop new therapies, are urgently needed. Three dimensional (3D) in vitro models are offering intriguing alternatives to conventional in vitro and in vivo models. Methods: We generated a 3D in vitro model using a primary HNSCC cell line derived from larynx tumor specimen and a commercial cell line (FaDu). Patient‐derived cells underwent characterization using Whole Exome Sequencing, WES. We investigated the morphology and protein expression of two cell lines, a FaDu and a primary cell line, on a 2D and 3D. We used spongy scaffolds of PVA/Gelatin (G) (70/30 w/w), which were then frozen and lyophilized. Primary HNSCCs, and FaDu cells were seeded at 500 × 103 cells/scaffold. These constructions were cultivated for 15 days and then tested at different day and 15 days. The metabolic assay was run during the culturing period. The constructions were processed for histology at each time point. Results: WES show 10‐20 mutations. Histology and immunohistochemistry showed that neoplastic cells in the scaffold/cell system resembled quite well the HNSCC. In 3D models, neoplastic cell morphology is much closer to the native tumor than in 2D models, as compared to the patient's HNSCC histologic specimen. We verified the expression of vimentin, P63, TP53, pancytokeratin, PD‐L1. Conclusion/Significance: The PVA/G sponges mimick the epithelial squamous environment, allowing the study of the mechanisms of development of HNSCC.