Department of Biomedical Engineering, University of Illinois Chicago, Chicago, 851 S. Morgan Street, 218 SEO, Chicago, IL 60607, USA.
Department of Surgery, University of Illinois Chicago, Chicago, IL 60612, USA.
Lab Chip. 2024 Mar 26;24(7):2025-2038. doi: 10.1039/d3lc01044a.
Patient-derived organoids (PDOs) serve as invaluable 3D tumor models, retaining the histological complexity and genetic heterogeneity found in primary tumors. However, the limitation of small sample volumes and the lack of tailored platforms have hindered the research using PDOs. Within the tumor microenvironment, cancer-associated fibroblasts play a pivotal role in influencing drug sensitivity. In this study, we introduce an agarose microwell platform designed for PDO-based tumor and tumor microenvironment models, enabling rapid drug screening and resistance studies with small sample volumes. These microwells, constructed using 3D printing molds, feature a U-shaped bottom and 200 μm diameter. We successfully generated co-culture spheroids of non-small cell lung carcinoma (NSCLC) cells, including NCI-H358 or A549, and NSCLC PDOs F231 or F671 with fibroblast cell line, WI-38. Our results demonstrate the production of uniformly-sized spheroids (coefficient of variation <30%), high viability (>80% after 1 week), and fibroblast-induced drug resistance. The PDOs maintained their viability (>81% after 2 weeks) and continued to proliferate. Notably, when exposed to adagrasib, a KRAS inhibitor, we observed reduced cytotoxicity in KRAS-mutant spheroids when co-cultured with fibroblasts or their supernatant. The fibroblast supernatant sustained proliferative signals in tumor models. Taking into account the physical features, viability, and drug resistance acquired through supernatants from the fibroblasts, our platform emerges as a suitable platform for tumor modeling and the evaluation of drug efficacy using patient-derived tissues.
患者来源的类器官（PDO）作为非常有价值的 3D 肿瘤模型，保留了原发性肿瘤中发现的组织学复杂性和遗传异质性。然而，小样本量的限制和缺乏定制平台限制了 PDO 的研究。在肿瘤微环境中，癌症相关成纤维细胞在影响药物敏感性方面起着关键作用。在这项研究中，我们引入了一种琼脂糖微井平台，用于基于 PDO 的肿瘤和肿瘤微环境模型，能够以小样本量进行快速药物筛选和耐药性研究。这些微井使用 3D 打印模具构建，具有 U 形底部和 200μm 直径。我们成功地生成了非小细胞肺癌（NSCLC）细胞的共培养球体，包括 NCI-H358 或 A549，以及 NSCLC PDO F231 或 F671 与成纤维细胞系 WI-38。我们的结果表明产生了均匀大小的球体（变异系数 <30%）、高活力（>80%在 1 周后）和成纤维细胞诱导的耐药性。PDO 保持活力（>81%在 2 周后）并继续增殖。值得注意的是，当暴露于 KRAS 抑制剂阿达格拉西布时，我们观察到与成纤维细胞或其上清液共培养时，KRAS 突变球体的细胞毒性降低。成纤维细胞上清液维持肿瘤模型中的增殖信号。考虑到物理特征、活力和通过成纤维细胞上清液获得的耐药性，我们的平台成为使用患者来源组织进行肿瘤建模和评估药物疗效的合适平台。
