Miller Paula G, Huang Emina, Fisher Robert, Shuler Michael L
Department of Biomedical Engineering, Cornell University, Ithaca, New York, USA.
Department of Surgery, UT Southwestern Medical Center, Dallas, Texas, USA.
Biotechnol Bioeng. 2025 Mar;122(3):481-494. doi: 10.1002/bit.28890. Epub 2024 Nov 25.
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.
我们描述了一种新型装置，用于在人体模型中模拟结肠癌细胞向肝脏的转移。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的五个步骤：在结肠中的侵袭、进入血流的血管内渗、全身运输、进入肝脏的血管外渗以及在肝脏中的定植。结肠模拟物使用接种在膜两侧的成熟正常结肠上皮类器官细胞（NL）和人脐静脉内皮细胞（HUVEC）。为了更好地模拟结肠结构，膜的NL侧暴露于空气中以建立气液界面。肝脏模拟物由接种在基质胶中膜两侧的人肝窦内皮细胞（HHSEC）和肝上皮细胞（HepG2 C3A）组成。在系统组装前，将来自类器官的标记结肠癌细胞/细胞团（CA）引入同一位患者已建立的正常结肠上皮细胞（NL）层中，或者将来自同一位患者的NL类器官和荧光标记的CA类器官按10:1的NL:CA比例制备，并在系统组装前一起建立。在该系统中细胞活力大于85%。我们证明，在5天的运行过程中，转移级联的五个步骤得以重现。这种新型装置允许针对选定变量在体外评估转移能力（通过使用ImageJ测量每个装置中标记区域的百分比）。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）。首次，这个新的体外系统在单个装置中模拟了转移级联的所有五个步骤，并提供了一个新装置，可在仅5天内以人体模型探测和观察转移过程。快速观察是由于在结肠中使用了高浓度的癌细胞（例如10%）且不存在免疫系统。我们的装置使得探究转移各步骤的方面以及各步骤之间的相互作用成为可能。
