Breier Joseph M, Radio Nicholas M, Mundy William R, Shafer Timothy J
The Curriculum in Toxicology, UNC School of Medicine, The University of North Carolina at Chapel Hill, Chapel Hill, North Carolina 27599, USA.
Toxicol Sci. 2008 Sep;105(1):119-33. doi: 10.1093/toxsci/kfn115. Epub 2008 Jun 11.
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 approximately 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.
公众普遍担忧，大多数商业化学品尚未针对其导致发育性神经毒性的可能性进行评估。尽管按照当前的发育性神经毒性指南，每年会对几种化学品进行评估，但时间、资源和动物实验的限制使得无法使用这种方法对大量化学品进行测试。因此，开发体外方法以筛选化学品对发育中的人类神经系统的潜在危害的动力越来越大。作为朝着这个目标迈出的第一步，本研究评估了一种自动化的高通量方法，该方法使用人神经祖细胞（hNPC）系ReNcell CX细胞来筛选化学品对细胞增殖和活力的影响。ReNcell CX细胞大约每36小时增殖一倍，并表达神经祖细胞标志物巢蛋白和SOX2。使用已知的抗增殖化合物对细胞增殖（5-溴-2'-脱氧尿苷掺入）和活力（碘化丙啶排除）的高通量测定方法进行了优化和测试。使用一组包含8种已知会导致发育性神经毒性的化合物和8种可能无毒的化合物的化合物集，进一步评估了这种体外筛选方法的实用性。8种发育性神经毒性物质中有6种显著抑制了ReNcell CX细胞的增殖和/或活力，而8种无毒化学品中有2种仅产生了最小的影响。这些结果表明，可以通过使用hNPC的高通量方法评估化学品对细胞增殖和活力的影响。作为筛选化合物对神经系统发育潜在影响的策略的一部分，进一步开发这种方法是有必要的。
