PO.TB10.18 · 肿瘤生物学
利用微生理系统构建血管化三维类肿瘤模型
Development of vascularized 3D tumoroid models using microphysiological systems
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
随着该领域寻求以人类相关的预测模型取代动物实验，新方法学（New Approach Methodologies，NAMs）在药物发现和毒理学中的应用正在加速。微生理系统（MPS）通过实现生理相关、可重复的体外模型引领着这一转变。在此，我们展示了利用AIM Biotech的VasQ Kit和organiX™平台以及Thermo Fisher Scientific的患者来源OncoPro™类肿瘤细胞系构建血管化类肿瘤模型。这些模型将针对复杂三维（3D）共培养优化的微流控系统与生理相关的类肿瘤（癌症类器官）系相结合。VasQ Kit中经过预验证的内皮细胞和成纤维细胞与OncoPro类肿瘤模型在水凝胶基质中共培养，以生成可灌注的血管网络。来源于结直肠癌（HuCo1044-GFP）和肺癌（HuLu051421）的类肿瘤被整合到organiX器件中，以评估肿瘤-血管相互作用。使用Texas Red-葡聚糖、抗CD31以及CellEvent™ Caspase-3/7试剂进行荧光和共聚焦成像，以可视化类肿瘤内的灌注、血管结构和凋亡。
可灌注的血管网络在患者来源类肿瘤周围可重复地形成，重现了肿瘤微环境（TME）的关键特征。共聚焦成像揭示了活跃的血管-类肿瘤界面，内皮网络穿透水凝胶基质并包绕肿瘤团块。凋亡标志物的定量以及免疫或基质细胞区室的空间分析证明了该模型评估药物反应和细胞浸润的能力。
该方法建立了一种标准化的、人类相关的血管化类肿瘤MPS模型，可用于肿瘤生物学、免疫细胞迁移和治疗疗效的机制研究。这一整合方法支持向NAMs的更广泛转变，并为免疫肿瘤学和精准医学研究提供了一个具有转化相关性的平台。
查看英文原文 English abstract
The adoption of New Approach Methodologies (NAMs) in drug discovery and toxicology is accelerating as the field seeks human-relevant predictive models to replace animal testing. Microphysiological systems (MPS) are leading this transition by enabling physiologically relevant, reproducible in vitro models. Here, we demonstrate construction of vascularized tumoroid models using AIM Biotech's VasQ Kit and organiX™ platform and Thermo Fisher Scientific patient-derived OncoPro™ Tumoroid Cell Lines. These models combine a microfluidic system optimized for complex three-dimensional (3D) co-cultures with physiologically-relevant tumoroid (cancer organoid) lines. Endothelial cells and fibroblasts, which came pre-validated in the VasQ Kit, were co-cultured with OncoPro tumoroid models in hydrogel matrices to generate perfusable vascular networks. Tumoroids derived from colorectal (HuCo1044-GFP) and lung (HuLu051421) cancers were integrated into organiX devices to assess tumor-vasculature interactions. Fluorescent and confocal imaging with Texas Red-dextran, anti-CD31, and CellEvent™ Caspase-3/7 reagents were used to visualize perfusion, vascular architecture, and apoptosis within tumoroids.
Perfusable vascular networks reproducibly formed around patient-derived tumoroids, recapitulating critical features of the tumor microenvironment (TME). Confocal imaging revealed active vasculature-tumoroid interfaces, with endothelial networks penetrating hydrogel matrices and surrounding tumor masses. Quantification of apoptotic markers and spatial analysis of immune or stromal cell compartments demonstrated the model's capability for assessing drug response and cellular infiltration.
This method establishes a standardized and human-relevant MPS model for vascularized tumoroids, enabling mechanistic studies of tumor biology, immune cell trafficking, and therapeutic efficacy. This integrated approach supports the broader transition toward NAMs and offers a translationally relevant platform for immuno-oncology and precision medicine research.
利益披露 Disclosure
G. Sahni,
AIM Biotech Pte Ltd Employment.
H. Loo,
AIM Biotech Pte Ltd Employment.
C. D. Paul,
Thermo Fisher Scientific Employment, Stock.
M. R. Dallas,
Thermo Fisher Scientific Employment.
S. Lim,
AIM Biotech Pte Ltd Employment.
K. Mun,
AIM Biotech Pte Ltd Employment.