摘要
目的建立体外胚胎干细胞神经发育毒性评价模型,并对其有效性进行验证。方法胚胎干细胞(ES)体外悬滴、悬浮培养后,经5×10-7mol/L视黄酸诱导培养分化获得神经细胞,利用RT-PCR方法分析不同浓度青霉素G、苯妥英钠(DPH)和氟尿嘧啶(5-FU)对ES细胞定向分化为特异表达巢蛋白基因的神经细胞的影响,计算其对ES细胞神经定向分化能力的抑制作用。同时应用MTT法测定5-FU、DPH和青霉素G对ES和BALB/c细胞活性的抑制作用,结合分化抑制作用结果评价受试物的发育毒性。结果青霉素G、苯妥英钠和氟尿嘧啶的ID50D3巢蛋白分别为0.017、49.4和1139μg/ml,其发育毒性的判定依次无、弱和强。结论建立的胚胎干细胞神经发育毒性评价模型能够正确判定氟尿嘧啶、苯妥英钠和青霉素G的神经发育毒性。
Objective To establish embryonic stem cell（EST） for evaluation of developmental neurontoxicity and test the validation of EST.Methods RT-PCR method was used to study the influence of fluorouracil（5-FU）,diphenylhydantoin（DPH） and Penicillin G at the different concentrations on neuron cells with nestin expression that differentiate from embryonic stem cells（ES）.With the results of cells viability（ES and BALB /c）assessed by MTT,the developmental neurotoxicty characteristics of 5-FU,DPH and Penicillin G may be identified clearly.Results The ID50 D3 nestin concentration of three embryotoxicants were 0.017,49.4and 1139μg /ml.Penicillin G,DPH and 5-FU were discriminated as none-,weak-and strong-developmental neurotoxicity.Conclusion Test compounds among three classes were discriminated correctly.The EST on developmental neurontoxicity could be applied to the safety evaluation in vitro.
出处
《卫生研究》
CAS
CSCD
北大核心
2010年第6期685-688,共4页
Journal of Hygiene Research
基金
"十一五"国家科技支撑计划重大项目(No.2006BAK02A07)
农业部科技行业科技项目(No.200903054-06)
中国疾病预防控制中心青年科研基金资助项目(No.2010A206)
关键词
胚胎干细胞
神经发育毒性
embryonic stem cells
developmental neurotoxicity
validation
