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P57. Defining success parameters for appendiceal cancer organoids: the first comprehensive analysis of 223 specimens from 78 patients

Thursday, March 5, 2026
2:53 PM - 3:00 PM MST
Location: 301A
Introduction: Patient-derived tumor organoids represent a transformative tool for precision oncology, yet their application to appendiceal cancers remains limited due to perceived biological challenges, particularly mucin content. We systematically evaluated factors influencing organoid generation success in the largest cohort of appendiceal tumor specimens analyzed to date to identify clinical and pre-analytic predictors of success.


Methods: We analyzed 223 tumor specimens from 78 patients with appendiceal cancers undergoing cytoreductive surgery with hyperthermic intraperitoneal chemotherapy. Specimens underwent standardized enzymatic digestion and organoid culture in hydrogel matrices for 14 days. Viability and yield were quantified via live/dead fluorescence and luminescence-based assays. Over 30 clinical, pathological, and processing variables were evaluated by univariate and multivariate regression to identify predictors of successful organoid generation (tissue viability ≥ 74%, viable cell density ≥ 6,000 cells/mg).


Results: Of 223 specimens, 80.3% underwent same-day dissociation and 76.2% contained mucinous stroma. Median total yield was 7.6M cells (IQR 2.0–15.7 M), viability 75.9%, and viable count 5.6M. Median viable cell density was 3,073 cells/mg (IQR 1,362–5,951). Prior surgery independently predicted reduced organoid viability (OR = 0.258, p = 0.0019), a 74% decrease in odds of high viability. Larger cell diameter also predicted lower viability (OR = 0.396, p = 0.0005). Multi-site sampling improved organoid viability (Δ +0.649, p = 0.0088), while post-processing mass < 1 g predicted poorer outcomes (p = 0.0053). Neoadjuvant therapy reduced performance (Δ −0.481, p = 0.0393). Same-day processing trended toward benefit (OR = 2.33, p = 0.093). Mucin presence was not detrimental (p = 0.087), challenging the assumption that mucinous tumors are unsuitable for organoid culture.


Conclusions: Successful organoid generation from appendiceal cancers is significantly influenced by modifiable clinical and technical factors rather than mucin content. Key actionable predictors include avoiding previously operated tissue sites, implementing multi-site sampling, ensuring adequate tissue mass (greater than 1 gram), and prioritizing same-day processing, providing an evidence-based framework for optimizing organoid protocols in mucin-rich tumors.

Learning Objectives:

  • Identify clinical and technical factors that predict appendiceal cancer organoid success and apply evidence-based strategies to optimize specimen procurement and processing protocols.
  • Recognize how modifiable pre-analytic variables—such as tissue mass, timing, and sampling strategy—influence organoid viability and apply these insights to improve reproducibility in translational research.
  • Understand how standardized processing and data-driven modeling can enhance reproducibility and scalability of organoid-based oncology research.