Related Experiment Video
Updated: Jul 10, 2026
Establishment of a Co-culture System of Patient-Derived Colorectal Tumor Organoids and Tumor-Infiltrating Lymphocytes (TILs)
Published on: June 27, 2025
Development and clinical application of a primary cancer organoid-PBMC co-culture system for personalized
Xueping Wang1, Jinsui Li2, Xingping Wu1
1State Key Laboratory of Oncology in South China, Collaborative Innovation Center for Cancer Medicine, Guangdong Esophageal Cancer Institute, Guangdong Provincial Clinical Research Center for Cancer, Sun Yat-sen University Cancer Center, Guangzhou 510060, China.
Aim:
Many patients with non-small cell lung cancer (NSCLC) do not derive clinical benefit from immune checkpoint inhibitors (ICIs), and a reliable method for identifying potential responders is lacking. This study aims to establish an optimized patient-derived organoids (PDOs) culture system and develop a novel organoid-peripheral blood mononuclear cell (PBMC) co-culture platform to evaluate response to ICIs.
Methods:
We optimized culture conditions for patient-derived NSCLC organoids by integrating conditional reprogramming techniques. ICIs response was assessed with an organoid-PBMC co-culture system. The accuracy of this ex vivo system was validated by comparing its results with efficacy of ICI therapy in both an in vivo humanized mouse xenograft model and the corresponding patient's clinical response. Moreover, the genomic stability of the organoids during serial passaging was monitored through gene sequencing.
Results:
An optimal indirect co-culture system was established, culturing organoids with 25% Matrigel. This system consisted of 40 Gy-irradiated Swiss 3T3-J2 fibroblasts in an upper chamber and primary NSCLC cells in a lower chamber, using F-medium (DMEM/F-12 (3:1, v/v) + 10 µM Y-27632) supplemented with 0.5 µM A83-01. This method achieved an 85.59% (95/111) success rate. Using this platform, we evaluated PD-1 antibody efficacy with the organoid-PBMC co-culture system in 70 cases. For in vivo validation, a direct patient-derived organoid xenograft (PDOX) models were established in humanized mice reconstituted with the corresponding patient's PBMCs. Notably, the ex vivo results demonstrated a strong correlation with both the in vivo validation data and patients' actual clinical responses to ICIs, yielding a sensitivity of 100% and a specificity of 62.20%.
Conclusion:
We developed a novel, simple, rapid and reliable organoid-PBMC co-culture system for screening the response of NSCLC patients to ICI therapy.
Insights
A new organoid-peripheral blood mononuclear cell (PBMC) co-culture system accurately predicts non-small cell lung cancer (NSCLC) patient response to immune checkpoint inhibitors (ICIs). This reliable platform aids in identifying patients likely to benefit from ICI therapy.
Area of Science:
- Oncology
- Immunotherapy
- Biotechnology
Background:
- Many non-small cell lung cancer (NSCLC) patients do not respond to immune checkpoint inhibitors (ICIs).
- A predictive biomarker for ICI response in NSCLC is currently lacking.
- Patient-derived organoids (PDOs) offer a promising model for personalized medicine.
Purpose of the Study:
- To optimize a patient-derived organoid (PDO) culture system.
- To develop a novel organoid-peripheral blood mononuclear cell (PBMC) co-culture platform for evaluating ICI response.
- To validate the predictive accuracy of the ex vivo co-culture system.
Main Methods:
- Optimized NSCLC PDO culture using conditional reprogramming.
- Established an indirect organoid-PBMC co-culture system with specific media and feeder cells.
- Validated ex vivo results against in vivo xenograft models and patient clinical responses.
- Monitored genomic stability of organoids during serial passaging.
Main Results:
- Achieved an 85.59% success rate in establishing the organoid culture system.
- The organoid-PBMC co-culture system demonstrated 100% sensitivity and 62.20% specificity in predicting ICI response.
- Ex vivo predictions strongly correlated with in vivo and clinical outcomes.
- The system successfully evaluated PD-1 antibody efficacy in 70 cases.
Conclusions:
- Developed a novel, simple, rapid, and reliable organoid-PBMC co-culture system.
- This platform enables effective screening of NSCLC patient response to ICI therapy.
- The system holds potential for guiding personalized immunotherapy decisions in NSCLC.
