Protocol for generating a 3D hydrogel-based tumor resection model in vitro using pancreatic cancer cells

We present a protocol for the generation of two distinct in vitro tumor resection models to evaluate cellular responses at tumor margins in 3D. We describe steps for patterning pancreatic cancer cells (Panc-01) embedded in hydrogels using a custom 96-pin metal lid to create standardized resection cavities. In a second model, we detail procedures for refining the protocol to imitate narrow surgical resection margins. This protocol supports diverse treatment modalities and enables reproducible, high-throughput analysis of post-resection responses. For complete details on the use and execution of this protocol, please refer to Miebach et al.¹.