We describe a novel device to mimic the metastasis of cancer cells from the colon into the liver in a human model. The colon mimic is connected to the liver model by a gravity‐driven recirculating unidirectional flow of a blood surrogate and can mimic the five steps of the metastatic cascade: invasion in the colon, intravasation into the bloodstream, systemic transportation, extravasation into the liver, and colonization in the liver. The colon mimic uses established normal colon epithelial organoid cells (NL) and human umbilical vein endothelial cells (HUVEC) plated on opposite sides of a membrane. To better mimic the colon structure the NL side of the membrane is exposed to air to establish an air‐liquid interface. The liver mimic consists of human liver sinusoidal endothelial cells (HHSEC) and epithelial hepatic cells (HepG2 C3A) plated in Matrigel on opposite sides of a membrane. Labeled colorectal cancer cells/clusters (CA) from organoids are introduced into an established normal colon epithelial cell (NL) layer from the same patient before assembly of the system or alternatively NL organoids and fluorescently labeled CA organoids from the same patient were prepared as a ratio of 10:1 NL:CA and established together before assembly of the system. Cell viability is greater than 85% in this system. We demonstrate that over 5 days of operation that the five steps of the metastatic cascade are replicated. This novel device allows an in vitro estimate of metastatic capability (as measured by using percentages of the labeled areas per device through ImageJ) in response to selected variables. In this study, the metastatic capability depends on the source of cancer cells (e.g., the patient), the clumping of cancer cells, glucose concentration, and oxygen levels (hypoxia). For the first time, this new in vitro system mimics all five steps of the metastatic cascade in a single device and provides a new device to probe and observe the process of metastasis in a human‐based model in only 5 days. The rapid observation is due to the use of a high concentration of cancer cells in the colon (e.g. 10%) and the absence of the immune system. Our device makes it possible to probe aspects of each step of metastasis and interactions between steps.

中文翻译：

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开发微生理系统来模拟人类癌症从结肠到肝脏的转移


我们描述了一种新型装置，可以在人体模型中模拟癌细胞从结肠转移到肝脏的过程。结肠模拟物通过血液替代物的重力驱动循环单向流动与肝脏模型相连，并且可以模拟转移级联反应的五个步骤：侵袭结肠、渗入血流、全身运输、外渗进入肝脏和在肝脏定植。结肠模拟物使用已建立的正常结肠上皮类器官细胞 （NL） 和人脐静脉内皮细胞 （HUVEC） 电镀在膜的相对两侧。为了更好地模拟结肠结构，将膜的 NL 侧暴露在空气中以建立气液界面。肝脏模拟物由人肝窦内皮细胞 （HHSEC） 和上皮肝细胞 （HepG2 C3A） 组成，这些细胞接种在膜相对两侧的基质胶中。在系统组装之前，将来自类器官的标记的结直肠癌细胞/簇 （CA） 引入来自同一患者的已建立的正常结肠上皮细胞 （NL） 层，或者将来自同一患者的 NL 类器官和荧光标记的 CA 类器官制备为 10：1 NL：CA 的比例，并在系统组装前一起建立。该系统中的细胞活力大于 85%。我们证明，在 5 天的手术中，转移级联反应的五个步骤被复制。这种新型设备允许对所选变量的转移能力进行体外估计（通过使用通过 ImageJ 使用每个设备标记区域的百分比来测量）。在本研究中，转移能力取决于癌细胞的来源（例如患者）、癌细胞的聚集、葡萄糖浓度和氧气水平（缺氧）。 这种新的体外系统首次在单个设备中模拟了转移级联反应的所有五个步骤，并提供了一种新设备，可以在短短 5 天内在基于人体的模型中探测和观察转移过程。快速观察是由于结肠中使用了高浓度的癌细胞（例如 10%）和缺乏免疫系统。我们的设备可以探测转移的每个步骤的各个方面以及步骤之间的相互作用。