Interdisciplinary Faculty of Toxicology, College Station, TX, 77843, United States; Department of Veterinary Integrative Biosciences, Texas A&M University, College Station, TX, 77843, USA.
Departments of Molecular Biology and Biochemistry, USA.
Toxicology. 2020 Dec 1;445:152601. doi: 10.1016/j.tox.2020.152601. Epub 2020 Sep 24.
Angiogenesis is a complex process that is required for development and tissue regeneration and it may be affected by many pathological conditions. Chemicals and drugs can impact formation and maintenance of the vascular networks; these effects may be both desirable (e.g., anti-cancer drugs) or unwanted (e.g., side effects of drugs). A number of in vivo and in vitro models exist for studies of angiogenesis and endothelial cell function, including organ-on-a-chip microphysiological systems. An arrayed organ-on-a-chip platform on a 96-well plate footprint that incorporates perfused microvessels, with and without tumors, was recently developed and it was shown that survival of the surrounding tissue was dependent on delivery of nutrients through the vessels. Here we describe a technology transfer of this complex microphysiological model between laboratories and demonstrate that reproducibility and robustness of these tissue chip-enabled experiments depend primarily on the source of the endothelial cells. The model was highly reproducible between laboratories and was used to demonstrate the advantages of the perfusable vascular networks for drug safety evaluation. As a proof-of-concept, we tested Fluorouracil (1-1,000 μM), Vincristine (1-1,000 nM), and Sorafenib (0.1-100 μM), in the perfusable and non-perfusable micro-organs, and in a colon cancer-containing micro-tumor model. Tissue chip experiments were compared to the traditional monolayer cultures of endothelial or tumor cells. These studies showed that human in vitro vascularized micro-organ and micro-tumor models are reproducible organ-on-a-chip platforms for studies of anticancer drugs. The data from the 3D models confirmed advantages of the physiological environment as compared to 2D cell cultures. We demonstrated how these models can be translated into practice by verifying that the endothelial cell source and passage are critical elements for establishing a perfusable model.
血管生成是一个复杂的过程，对于组织发育和再生是必需的，它可能受到许多病理状况的影响。化学物质和药物会影响血管网络的形成和维持；这些影响可能是期望的（例如，抗癌药物），也可能是不期望的（例如，药物的副作用）。目前存在许多用于研究血管生成和内皮细胞功能的体内和体外模型，包括器官芯片微生理系统。最近开发了一种在 96 孔板足迹上排列的器官芯片平台，该平台包含灌注的微血管，有或没有肿瘤，并表明周围组织的存活取决于通过血管输送营养物质。在这里，我们描述了在实验室之间进行这种复杂的微生理模型的技术转移，并证明了这些组织芯片实验的重现性和稳健性主要取决于内皮细胞的来源。该模型在实验室之间具有高度重现性，并用于证明可灌注血管网络在药物安全性评估中的优势。作为概念验证，我们在可灌注和不可灌注的微器官以及含有结肠癌的微肿瘤模型中测试了氟尿嘧啶（1-1000 μM）、长春新碱（1-1000 nM）和索拉非尼（0.1-100 μM）。将组织芯片实验与内皮细胞或肿瘤细胞的传统单层培养进行了比较。这些研究表明，人类体外血管化微器官和微肿瘤模型是用于研究抗癌药物的可重现的器官芯片平台。与 2D 细胞培养相比，3D 模型的数据证实了生理环境的优势。我们通过验证内皮细胞来源和传代对于建立可灌注模型是关键因素，证明了如何将这些模型转化为实践。
