Nguyen Eric H, Dombroe Micah J, Fisk Debra L, Daly William T, Sorenson Christine M, Murphy William L, Sheibani Nader
Human Models for Analysis of Pathways Center, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin.
Department of Ophthalmology and Visual Sciences, University of Wisconsin School of Medicine and Public Health, Madison, Wisconsin.
Appl In Vitro Toxicol. 2019 Jun 1;5(2):92-110. doi: 10.1089/aivt.2018.0025. Epub 2019 Jun 17.
Human-induced pluripotent stem cells (iPSCs) represent a promising cell source for the construction of organotypic culture models for chemical toxicity screening and characterization. To characterize the effects of chemical exposure on the human neurovasculature, we constructed neurovascular unit (NVU) models consisting of endothelial cells (ECs) and astrocytes (ACs) derived from human-iPSCs, as well as human brain-derived pericytes (PCs). The cells were cocultured on synthetic poly(ethylene glycol) (PEG) hydrogels that guided the self-assembly of capillary-like vascular networks. High-content epifluorescence microscopy evaluated dose-dependent changes to multiple aspects of NVU morphology. Cultured vascular networks underwent quantifiable morphological changes when incubated with vascular disrupting chemicals. The activity of predicted vascular disrupting chemicals from a panel of 38 compounds (U.S. Environmental Protection Agency) was ranked based on morphological features detected in the NVU model. In addition, unique morphological neurovascular disruption signatures were detected per chemical. A comparison of PEG-based NVU and Matrigel-based NVU models found greater sensitivity and consistency in chemical detection by the PEG-based NVU models. We suspect that specific morphological changes may be used for discerning adverse outcome pathways initiated by chemical exposure and rapid mechanistic characterization of chemical exposure to neurovascular function. The use of human stem cell-derived vascular tissue and PEG hydrogels in the construction of NVU models leads to rapid detection of adverse chemical effects on neurovascular stability. The use of multiple cell types in coculture elucidates potential mechanisms of action by chemicals applied to the model.
人诱导多能干细胞（iPSC）是构建用于化学毒性筛选和表征的器官型培养模型的一种有前景的细胞来源。为了表征化学物质暴露对人类神经血管系统的影响，我们构建了神经血管单元（NVU）模型，该模型由源自人iPSC的内皮细胞（EC）、星形胶质细胞（AC）以及人脑来源的周细胞（PC）组成。这些细胞在引导类毛细血管状血管网络自组装的合成聚乙二醇（PEG）水凝胶上共培养。高内涵落射荧光显微镜评估了NVU形态多个方面的剂量依赖性变化。当与血管破坏化学物质一起孵育时，培养的血管网络发生了可量化的形态变化。根据在NVU模型中检测到的形态特征，对来自一组38种化合物（美国环境保护局）的预测血管破坏化学物质的活性进行了排名。此外，每种化学物质都检测到了独特的形态学神经血管破坏特征。基于PEG的NVU模型和基于基质胶的NVU模型的比较发现，基于PEG的NVU模型在化学物质检测方面具有更高的灵敏度和一致性。我们怀疑特定的形态变化可用于识别由化学物质暴露引发的不良结局途径以及对化学物质暴露于神经血管功能的快速机制表征。在NVU模型构建中使用人干细胞衍生的血管组织和PEG水凝胶可快速检测化学物质对神经血管稳定性的不良影响。共培养中使用多种细胞类型阐明了应用于该模型的化学物质的潜在作用机制。
