Endothelial Progenitor Cells for Assessing Toxicological Response Variability in

目前，毒理学风险评估缺乏对以下物质的大小和变异性的直接估计 人类对环境毒物的反应。具体地说，关于Low的影响的数据非常少 剂量毒物对人类发育过程，包括细胞分化的影响。那是因为 目前毒理学评估的基础，动物和永生化细胞系，不能充分地 模拟低水平毒物暴露对人类人口的影响。一种潜在的选择 检测平台应能够在大量筛查中纳入人类变异性 毒物以高通量的方式。为了建立这样的平台，我们建议使用人类捐赠者- 特定的细胞可以(A)以非侵入性的方式分离，(B)容易在培养中扩增，以及(C) 超低温保存后不会失去活力。我们认为人内皮祖细胞或 内皮细胞集落形成细胞(ECFC)是一组CD31+/CD34+多能细胞，发现于 流通，将满足这些要求。在体外，ECFC高度增殖，可以 分化为成熟的内皮细胞。这项研究的总体目标是评估 化学毒物对人脐静脉内皮细胞活性、增殖和分化的影响 不同的个体。具体地说，在这项建议中，我们将确定环境保护措施作为一种 高通量化学毒性测试平台。为此，我们选择了三个已知的 新生儿和成人外周血中发现低水平环境毒物：双酚A (双酚A)、全氟辛酸(全氟辛酸)和镉。这项拟议的研究是 开发一种利用供体特异性多能细胞的新颖、强大和全面的平台。 在本项目结束时，我们将演示基于ECFC的检测方法对高 环境毒物的吞吐量评价。基于ECFC的毒理学检测可以成为一种 人类遗传多样性功能评估和种群识别的宝贵工具 最容易受到化学危害的亚群。

Currently, toxicological risk assessment lacks direct estimation of the magnitude and variability of human responses to environmental toxicants. Specifically, there is very little data on the effects of low dose toxicants on human developmental processes, including cellular differentiation. That is because the current basis of toxicological assessment, animals and immortalized cell lines, cannot adequately model the impact of low level toxicant exposure on human population. A potential alternative detection platform should enable incorporation of human variability in screening of a large number of toxicants in a high-throughput manner. To establish such a platform, we suggest using human donor- specific cells that can be (a) isolated in a non-invasive manner, (b) easily expanded in culture, and (c) cryopreserved without loss of viability. We propose that human endothelial progenitor cells or endothelial colony forming cells (ECFCs), a population of CD31+/CD34+ pluripotent cells found in circulation, would fulfill these requirements. In vitro, ECFCs are highly proliferative and can differentiate into mature endothelial cells (ECs). The overall goal of this research is to evaluate the effect of chemical toxicants on viability, proliferation, and differentiation of ECFCs derived from different individuals. Specifically in this proposal, we will determine the feasibility of ECFCs as a platform for high-throughput chemical toxicity testing. For this purpose, we selected three known environmental toxicants found at low levels in neonatal and adult peripheral blood: bisphenol A (BPA), perfluorooctanoic acid (PFOA), and cadmium. The proposed study is the key step in the development of a novel, robust, and comprehensive platform utilizing donor-specific pluripotent cells. At the conclusion of this project, we will demonstrate the amenability of ECFC-based assays for high- throughput evaluation of environmental toxicants. ECFC-based toxicological assays can become an invaluable tool for functional assessment of human genetic diversity and identification of population subgroups most vulnerable to chemical hazards.