Background <p>Tubo-ovarian carcinoma, a leading cause of gynaecological-related mortality, holds substantial biological and clinical heterogeneity. Despite advancements in drug development, predicting therapeutic efficacy remains challenging, partly due to the limited availability of in vitro models that accurately replicate tumour behaviour. We present a concise overview of the intrahospital workflow for establishing patient-derived organoids and analyse the morphological and immunophenotypical features of high-grade serous carcinoma (HGSC), serous borderline tumour (SBT)/low-grade serous carcinoma (LGSC), and normal fallopian tube (FT) organoids.</p> Results <p>Samples were collected from patients undergoing surgery or paracentesis. Tissue underwent mechanical and enzymatical digestion. Resulting cell suspensions were resuspended in an extracellular matrix substitute for subsequent culture. Despite the low efficacy in establishing HGSC organoids (<i>n</i> = 1/7, 14%; 96 days, 11 passages), we successfully established two organoid lines of SBT/LGSC (<i>n</i> = 2/2, 100%; 65 days, 7 passages; 134 days, 16 passages) and normal FT (<i>n</i> = 2/2, 100%; 73 days, 10 passages; 58 days, 8 passages). HGSC organoids exhibited limited growth and mostly irregular structures, while preserving the p53 immunostaining pattern of the original tumour. SBT/LGSC and FT organoids maintained features of architectural complexity and faithfully recapitulated the original immunoprofile.</p> Conclusions <p>This study highlights the need for a multidisciplinary collaboration in both clinical and research settings to establish patient-derived organoids. It emphasises the pivotal contribution of pathologists in meticulous sampling and organoid characterisation. The integration of diverse expertise is essential for maximising the potential of organoids as preclinical tools, advancing our understanding of tubo-ovarian carcinoma, and ultimately improving patient outcomes.</p>