Recombinant spider silk proteins inspired by natural Araneus diadematus fibroins (ADF) are highly biocompatible, non-inflammatory and versatile processable. In general, materials made of the negatively charged, recombinant eADF4(C16) variant show low cell adhesion. However, a guided cell interaction is a crucial pre-requisite for successful application in tissue engineering and regenerative medicine. Therefore, different modification strategies have evolved to functionalize spider silk scaffolds for adjusting the cell-substrate interaction. In this context, specific cell adhesion could be achieved by using either varying processing methods or genetic modification strategies. Thereby, the introduction of differently shaped and sized topographical surface patterns on eADF4(C16) films led to selective cell-material interactions. Furthermore, site specific amino acid exchanges were used to generate the positively charged eADF4(κ16) or the uncharged eADF4(Ω16) variant, which were further functionalized with cell adhesive peptides, such as the integrin recognition sequence RGD, to enable specific cell attachment on films made thereof.