Development of a high-throughput screening assay for chemical effects on proliferation and viability of immortalized human neural progenitor cells
Development of a high-throughput screening assay for chemical effects on proliferation and viability of immortalized human neural progenitor cells
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DOI：
10.1093/toxsci/kfn115
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发表时间：
2008-09-01
影响因子：
3.8
通讯作者：
Shafer, Timothy J.
中科院分区：
文献类型：
--
作者：
Breier, Joseph M.;Radio, Nicholas M.;Shafer, Timothy J.
公众十分关注的是，大多数商业化学品尚未对其造成发育神经毒性的潜力进行评估。虽然根据目前的发育神经毒性指南每年评估几种化学品，但时间、资源和动物限制阻止了使用这种方法对大量化学品进行测试。因此，开发体外方法来筛选可能损害发育中的人类神经系统的化学物质的动机越来越强烈。作为实现这一目标的第一步，本研究评估了一种自动化的高通量方法，用于使用ReNcell CX细胞（一种人神经祖细胞（hNPC）系）筛选对增殖和活力的化学作用。ReNcell CX细胞在类似于36小时内倍增，并表达神经祖细胞标记物巢蛋白和SOX 2。使用已知的抗增殖化合物优化和测试细胞增殖（5-溴-2 '-脱氧尿苷掺入）和活力（碘化丙啶排除）的高通量测定。使用一组含有八种已知引起发育神经毒性的化合物和八种可能无毒的化合物进一步评价了这种体外筛选的效用。八种发育神经毒物中有六种显着抑制ReNcell CX细胞增殖和/或活力，而八种无毒化学物质中有两种仅引起最小的影响。这些结果表明，可以通过使用hNPC的高通量方法评估对细胞增殖和活力的化学作用。作为筛选化合物对神经系统发育的潜在影响的策略的一部分，进一步开发这种方法是必要的。
There is considerable public concern that the majority of commercial chemicals have not been evaluated for their potential to cause developmental neurotoxicity. Although several chemicals are assessed annually under the current developmental neurotoxicity guidelines, time, resource, and animal constraints prevent testing of large numbers of chemicals using this approach. Thus, incentive is mounting to develop in vitro methods to screen chemicals for their potential to harm the developing human nervous system. As an initial step toward this end, the present studies evaluated an automated, high-throughput method for screening chemical effects on proliferation and viability using ReNcell CX cells, a human neural progenitor cell (hNPC) line. ReNcell CX cells doubled in similar to 36 h and expressed the neural progenitor markers nestin and SOX2. High-throughput assays for cell proliferation (5-bromo-2'-deoxyuridine incorporation) and viability (propidium iodide exclusion) were optimized and tested using known antiproliferative compounds. The utility of this in vitro screen was evaluated further using a set of compounds containing eight known to cause developmental neurotoxicity and eight presumably nontoxic compounds. Six out of eight developmental neurotoxicants significantly inhibited ReNcell CX cell proliferation and/or viability, whereas two out of eight nontoxic chemicals caused only minimal effects. These results demonstrate that chemical effects on cell proliferation and viability can be assessed via high-throughput methods using hNPCs. Further development of this approach as part of a strategy to screen compounds for potential effects on nervous system development is warranted.