Engineering complex tissue-like scaffolds by two photon polymerization for testing cancer cell behavior in confined environments

Cancer cells behavior in confined spaces is strongly related to cell-specific morphological deformations, which further influence the migration mechanism during the cancer metastasis processes. Two photon polymerization (TPP) was applied to tailor tissue-like scaffolds with narrow confined pores in polymeric materials. We have fabricated 3D scaffolds in SU8 photoresist by TPP, with pore sizes down to sub-micrometer dimensions. Such 3D platforms were further coated with collagen for testing the invasiveness potential of melanoma cancer cells in confined spaces. Quantitative analysis of cell adhesion and invasiveness within the scaffolds were performed by fluorescent microscopy of nuclei, cytoskeleton and focal adhesion points. We found that cellular affinity given by cell nuclei surrounding the scaffolds was two times higher when a collagen coating is was applied. The focal adhesion anchoring around the borders or inside the scaffolds was found much denser when collagen is was applied, but much stronger when collagen is was not used.