Head and Neck Squamous cell carcinomas in 3D organotypic co-cultures (OTC)

Introduction HNSCC display a high heterogeneity in molecular properties, cellular infiltrates and interaction with the microenvironment. For detailed studies we developed a three-dimensional organotypic culture enabling a better understanding of tumor progression and the interaction of the tumor, its microenvironment and the infiltrating immune cells

Methods The three-dimensional organotypic co-culture model (OTC) is composed by scaffold-based dermal equivalents from human fibroblasts and viscose fibers co-cultivated with vital HNSCC explants with the size of 3x3x3 mm freshly after operative resection in 4.5 ml medium. It allows the maintenance of individual tumor structures and microenvironment texture. Following defined kinetics harvested samples were paraffin fixed. To visualize OTC progression over time, hematoxylin eosin and immunohistochemical staining then were performed. The immune cell status of this tumor model was investigated via immunohistochemistry targeting CD3, CD20, CD56 and CD68.

Results OTCs stayed vital for up to 28 days in cultivation. Interestingly tumors develop different expansive and invasive growth patterns which can be correlated with eg. the tumor cell budding present in the primary tumor. The primary tumor′s immune cell status was evaluated (i.e. hot, cold, excluded). The kinetics of the immune cells in the respective OTCs was surveilled and classified.

Conclusions  The scaffold-based culture system allows tumor growth in its original environment by maintaining the tumor cell heterogeneity and cellular infiltrates. OTCs serve as a model that allows investigation of tumor cell budding based on the analysis of epithelial-mesenchymal transition and the tumor microenvironment as crucial components of metastasis.

Publication History

Article published online:
12 May 2023