Feng Zeshuo, Li Yanjiao, Wu Ju, Yin Jiajun, Wang Ruoyu, Liang Shanshan
The Key Laboratory of biomarker high throughput screening and target translation of breast and gastrointestinal tumor, Affiliated Zhongshan Hospital of Dalian University.
Department of General Surgery, Affiliated Zhongshan Hospital of Dalian University.
J Vis Exp. 2025 Jun 27(220). doi: 10.3791/68346.
Colorectal cancer (CRC) is the third most common cancer worldwide. Tumor-infiltrating lymphocytes (TILs) have been identified as an important prognostic marker in CRC. The therapeutic application of TILs has already shown promising results in melanoma and cervical cancer. However, their use in CRC therapy remains in an exploratory phase. A suitable in vitro model to evaluate TIL efficacy is currently unavailable, hindering further advancements in this field. Patient-derived organoid (PDO) models, which closely retain the characteristics of the original tumor tissue and reflect inter-patient heterogeneity, provide an excellent platform for studying the interaction between CRC and TILs. In this study, a method is described to establish a patient-derived CRC organoid model from freshly resected tumor tissue, followed by isolation and expansion of TILs. This system allows co-culture of CRC organoids and TILs, enabling the assessment of TIL-mediated cytotoxicity and immune responses. By analyzing TIL killing efficacy on organoids, the potential outcomes of TIL-based immunotherapy for personalized CRC treatment can be predicted. Moreover, further engineering of TILs may enhance their anti-tumor efficacy, offering a promising strategy for developing more effective cellular therapies. This PDO-TIL co-culture model provides a powerful tool for preclinical evaluation of TIL therapies and personalized treatment strategies in CRC.
结直肠癌（CRC）是全球第三大常见癌症。肿瘤浸润淋巴细胞（TILs）已被确定为CRC的一种重要预后标志物。TILs的治疗应用在黑色素瘤和宫颈癌中已显示出有前景的结果。然而，它们在CRC治疗中的应用仍处于探索阶段。目前尚无合适的体外模型来评估TILs的疗效，这阻碍了该领域的进一步发展。患者来源的类器官（PDO）模型紧密保留了原始肿瘤组织的特征并反映了患者间的异质性，为研究CRC与TILs之间的相互作用提供了一个极佳的平台。在本研究中，描述了一种从新鲜切除的肿瘤组织建立患者来源的CRC类器官模型的方法，随后进行TILs的分离和扩增。该系统允许CRC类器官与TILs共培养，从而能够评估TIL介导的细胞毒性和免疫反应。通过分析TILs对类器官的杀伤效果，可以预测基于TILs的免疫疗法用于个性化CRC治疗的潜在结果。此外，对TILs的进一步工程改造可能会增强其抗肿瘤疗效，为开发更有效的细胞疗法提供了一种有前景的策略。这种PDO-TIL共培养模型为CRC中TIL疗法的临床前评估和个性化治疗策略提供了一个强大的工具。
