Multi-Component Microscaffold With 3D Spatially Defined Proteinaceous Environment

In this paper, we present a multicomponent microenvironment consisting of proteinaceous networks with submicron-sized features optionally embedded into a photoresist microscaffold. By two-photon direct laser writing, free-standing 3D proteinaceous microstructures were fabricated for cell culture application, demonstrated with NIH/3T3 fibroblast cells. A Young's modulus of megapascal-order contributes to the challenge of structural sustainability of the proteinaceous microstructures for experiments as well as sequential fabrication steps. We propose to embed proteinaceous networks into a mechanically robust photoresist microscaffold. We investigate the limits of this 3D microfabrication of embedded proteinaceous networks and demonstrate the embedment of two different proteinaceous networks within one microscaffold. Performing cell culture of PC12 cells, we observe cell adhesion and cell motility on embedded proteinaceous networks of collagen type-IV mixed with bovine serum albumin into a photoresist microscaffold. The ability to structure proteinaceous elements for 3D spatial control of microenvironment might be a key feature in cell culture to decouple environmental cues to control cellular behavior.