Recent advances in anticancer therapy have shown dramatic improvements in clinical outcomes, and adoptive cell therapy has emerged as a type of immunotherapy that can modulate immune responses by transferring engineered immune cells. However, a small percentage of responders and their toxicity remain as challenges. Three-dimensional (3D) in vitro models of the tumor microenvironment (TME) have the potential to provide a platform for assessing and predicting responses to therapy. This paper describes an in vitro 3D tumor model that incorporates clusters of colorectal cancer (CRC) cells around perfusable vascular networks to validate immune-cell-mediated cytotoxicity against cancer cells. The platform is based on an injection-molded 3D co-culture model and composed of 28 microwells where separate identical vascularized cancer models can be formed. It allows robust hydrogel patterning for 3D culture that enables high-throughput experimentation. The uniformity of the devices resulted in reproducible experiments that allowed 10× more experiments to be performed when compared to conventional polydimethylsiloxane (PDMS)-based microfluidic devices. To demonstrate its capability, primary natural killer (NK) cells were introduced into the vascularized tumor network, and their activities were monitored using live-cell imaging. Extravasation, migration, and cytotoxic activity against six types of CRC cell lines were tested and compared. The consensus molecular subtypes (CMS) of CRC with distinct immune responses resulted in the highest NK cell cytotoxicity against CMS1 cancer cells. These results show the potential of our vascularized tumor model for understanding various steps involved in the immune response for the assessment of adoptive cell therapy.

抗癌治疗的最新进展显示出临床结果的显着改善，并且过继细胞疗法已成为一种免疫疗法，可以通过转移工程免疫细胞来调节免疫反应。然而，一小部分响应者及其毒性仍然是挑战。三维（3D）体外肿瘤微环境 (TME) 模型有可能为评估和预测治疗反应提供一个平台。本文描述了一个体外3D 肿瘤模型，在可灌注血管网络周围包含结直肠癌 (CRC) 细胞簇，以验证免疫细胞介导的对癌细胞的细胞毒性。该平台基于注射成型的 3D 共培养模型，由 28 个微孔组成，其中可以形成单独的相同血管化癌症模型。它可以为 3D 培养提供强大的水凝胶图案，从而实现高通量实验。与传统的基于聚二甲基硅氧烷 (PDMS) 的微流体装置相比，这些装置的均匀性导致了可重复的实验，可以进行 10 倍以上的实验。为了证明其能力，将原代自然杀伤 (NK) 细胞引入血管化肿瘤网络，并使用活细胞成像监测它们的活动。外渗、迁移、对六种类型的 CRC 细胞系的细胞毒活性进行了测试和比较。具有不同免疫反应的结直肠癌共有分子亚型 (CMS) 导致 NK 细胞对 CMS1 癌细胞的最高细胞毒性。这些结果显示了我们的血管化肿瘤模型在了解免疫反应中涉及的各个步骤以评估过继细胞治疗方面的潜力。