DEVELOPMENT OF AN IN VITRO MODEL FOR STUDYING PERINEURAL INVASION OF HEAD AND NECK SQUAMOUS CELL CARCINOMA

METHODS AND MATERIALS Abstract: Objective: To develop an in vitro model of perineural invasion of head and neck squamous cell carcinoma (HNSCCa) Study Design: Qualitative in vitro model Methods: Murine dorsal root ganglia (DRG) were extracted and embedded in Matrigel. FaDu HNSCCa cells were co-cultured with the DRG and multiple imaging methods were utilized to visualize the neurite and cell interactions. DRG were also dissociated into a sensory neuronal culture and co-cultured with FaDu cells and similarly imaged. Time- delayed images were recorded starting at 24 hours after co-culture. Two- and three-dimensional imaging techniques are also being utilized with GFP- and RFP-labeled FaDu cells. Results: DRG when embedded in Matrigel developed neurites that interacted with co-cultured FaDu cells with the cells growing along the neurites towards the DRG. Dissociated cells also interacted with co-cultured FaDu cells. Conclusions: This study presents a model of perineural invasion which has not been previously described using an HNSCCa cell line. Both in vitro methods have demonstrated reproducible perineural invasion of FaDu cells when co-cultured with either intact murine DRG or dissociated neuronal cells isolated from DRG. Ongoing studies are further quantifying the rate of perineural invasion as well as elucidating regulators and modulators of the neuron-cell interactions. The models are also being adapted to study chemotaxis in vitro.