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

## TL;DR

This paper describes a method to create 3D tumor models in hydrogels to study cellular responses at tumor margins after simulated surgery.

## Contribution

A novel protocol for generating standardized 3D tumor resection models using hydrogels and pancreatic cancer cells is introduced.

## Key findings

- A custom 96-pin metal lid is used to pattern pancreatic cancer cells in hydrogels for standardized resection cavities.
- The protocol allows for high-throughput analysis of post-resection cellular responses using various treatment modalities.
- The method supports reproducible and automated margin-specific analysis through viability assays and live imaging.

## Abstract

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

•Protocol for generating standardized 3D tumor resection cavities in hydrogel•Instructions for embedding tumor cells and creating defined resection margins•Guidance on applying chemical, oxidative, or physical post-resection treatments•Steps for viability assays and live imaging with automated margin-specific analysis

Protocol for generating standardized 3D tumor resection cavities in hydrogel

Instructions for embedding tumor cells and creating defined resection margins

Guidance on applying chemical, oxidative, or physical post-resection treatments

Steps for viability assays and live imaging with automated margin-specific analysis

Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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.