Tumor organoids are three-dimensional (3D) ex vivo tumor models that recapitulatethe biological key features of the original primary tumor tissues. Patient-derivedtumor organoids have been used in translational cancer research and can beapplied to assess treatment sensitivity and resistance, cell-cell interactions, and tumorcell interactions with the tumor microenvironment. Tumor organoids are complexculture systems that require advanced cell culture techniques and culture mediawith specific growth factor cocktails and a biological basement membrane thatmimics the extracellular environment. The ability to establish primary tumor cultureshighly depends on the tissue of origin, the cellularity, and the clinical featuresof the tumor, such as the tumor grade. Furthermore, tissue sample collection,material quality and quantity, as well as correct biobanking and storage are crucialelements of this procedure. The technical capabilities of the laboratory are also crucialfactors to consider. Here, we report a validated SOP/protocol that is technically andeconomically feasible for the culture of ex vivo tumor organoids from fresh tissuesamples of pancreatic adenocarcinoma origin, either from fresh primary resectedpatient donor tissue or patient-derived xenografts (PDX). The technique describedherein can be performed in laboratories with basic tissue culture and mouse facilitiesand is tailored for wide application in the translational oncology field.
