Background/Purpose: Conventional three-dimensional in vitro tumor models often fail to fully capture the complexity of the tumor microenvironment, particularly the diverse populations of cancer-associated fibroblasts that contribute to poor prognosis and treatment resistance. The purpose of this study is to develop a patient-specific gastric cancer assembloid model that integrates tumor epithelial cells with matched stromal cell subtypes, each derived using tailored growth media to enhance cancer preclinical research and advance personalized therapeutic strategies.Methods: Tumor tissue was dissociated, and cells expanded in media for organoids, mesenchymal stem cells, fibroblasts, or endothelial cells. The resulting tumor-derived subpopulations were co-cultured in an optimized assembloid medium supporting each cell type’s growth. Biomarker expression was assessed by immunofluorescence staining, and transcriptomic profiles were analyzed by RNA sequencing. Drug responsiveness was evaluated using cell viability assays following treatment with various therapeutic agents.Results: The optimized co-culture conditions yielded assembloids that closely mimicked the cellular heterogeneity of primary tumors, confirmed by the expression of epithelial and stromal markers. Compared to monocultures, the assembloids showed higher expression of inflammatory cytokines, extracellular matrix remodeling factors, and tumor progression-related genes across different organoids and stromal ratios. Drug screening revealed patient- and drug-specific variability. While some drugs were effective in both organoid and assembloid models, others lost efficacy in the assembloids, highlighting the critical role of stromal components in modulating drug responses.Conclusions: This assembloid system offers a robust platform to study tumor–stroma interactions, identify resistance mechanisms, and accelerate drug discovery and personalized therapeutic strategies for gastric cancer.
背景/目的：传统的三维体外肿瘤模型往往难以全面捕捉肿瘤微环境的复杂性，尤其是那些与不良预后和治疗抵抗相关的多种癌症相关成纤维细胞亚群。本研究旨在开发一种患者特异性胃癌组装体模型，该模型将肿瘤上皮细胞与匹配的基质细胞亚型相结合，每种细胞均通过定制化培养基进行扩增，以增强癌症临床前研究并推进个体化治疗策略。 方法：将肿瘤组织解离后，分别在类器官、间充质干细胞、成纤维细胞或内皮细胞专用培养基中进行细胞扩增。将获得的肿瘤来源细胞亚群在优化的组装体培养基中共培养，该培养基支持各类细胞的生长。通过免疫荧光染色评估生物标志物表达，并利用RNA测序分析转录组特征。使用多种治疗药物处理后，通过细胞活力测定评估药物反应性。 结果：优化的共培养条件成功构建出能高度模拟原发肿瘤细胞异质性的组装体，上皮及基质标志物的表达验证了这一点。与单一培养相比，组装体在不同类器官与基质比例条件下均表现出更高的炎症细胞因子、细胞外基质重塑因子及肿瘤进展相关基因的表达水平。药物筛选结果显示存在患者特异性及药物特异性的反应差异：部分药物在类器官和组装体模型中均有效，而另一些药物在组装体中疗效丧失，这凸显了基质成分在调节药物反应中的关键作用。 结论：该组装体系统为研究肿瘤-基质相互作用、识别耐药机制以及加速胃癌药物研发和个体化治疗策略提供了可靠平台。
肿瘤(癌症)患者之家