Protocol for the establishment and characterization of lung tumoroids from surgical resection without the use of Matrigel.

Organoids are vital for studying cancer mechanisms and treatment resistance, but their growth usually depends on costly, animal-derived Matrigel. Here, we present a protocol for expanding and maintaining lung-derived organoids without the need for Matrigel. We describe steps for establishing lung cancer organoids, staining organoids for immunohistochemistry, and cultivating them with ClinoStar. By eliminating dependence on Matrigel and its associated costs, this protocol enhances accessibility and scalability, making organoid-based research more feasible for cancer studies and therapeutic development.