以ITSFn诱导法为基础的神经发育毒性评价模型的建立及其有效性研究  

Study the developmental neurontoxicity evaluation model by ITSFn protocol and To verify its validity

作　　者：刘海波 支媛 冯永全 戴抒豪[2] 于洲 

机构地区：[1]国家食品安全风险评估中心,北京100021 [2]徐州医学院公共卫生学院,江苏徐州221004

出　　处：《中国食品卫生杂志》2013年第1期6-11,共6页Chinese Journal of Food Hygiene

基　　金：国家自然科学基金青年基金项目(81102134);农业部科技行业科技项目<农药风险评估综合配套技术研究>(200903054-06)

摘　　要：目的建立体外以胰岛素-转铁蛋白-硒-纤维连接蛋白(ITSFn)培养法为基础的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,完善神经发育毒性评价体系。方法胚胎干细胞悬滴、悬浮培养后,加入胰岛素、转铁蛋白、硒、纤维连接蛋白诱导神经细胞分化,结合细胞毒性MTT法建立神经发育毒性体外评价模型,并检测不同浓度青霉素G、苯妥英钠和氟尿嘧啶作用条件下其对胚胎干细胞神经分化能力的影响,判断受试物的神经发育毒性。结果青霉素G、苯妥英钠和氟尿嘧啶的ID50D3 nestin分别为0.017μg/ml、49.40μg/ml、1 139μg/ml,ID50D3EN-1分别0.031μg/ml、25.63μg/ml、1 762μg/ml,其发育毒性的判定均依次为无、弱和强。结论以ITSFn诱导法为基础建立的体外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的发育毒性,可用于体外神经发育毒性的筛选和评价。Objective Establishment of in vitro to insulin and transferrin-se-fibronectin （ITSFn） training method based on embryonic stem cells neural developmental toxicity evaluation model, and its effectiveness is verified, the refinement of neural development toxicity evaluation system. Methods embryonic stem ceils suspended drops, suspension culture, insulin, transferrin, adding selenium, fibronectin induces neuronal differentiation, in combination with cytotoxic MTT law to build the evaluation model of neurodevelopmental toxicity in vitro, and detection of different concentrations of penicillin G, phenytoin and 5 - fluorouracil effect under the condition of embryonic stem cell neural differentiation, ability, judgment the neurodevelopmental toxicity. Results penicillin G, phenytoin and 5-fluorouracil ID50D3 nestin were 0. 017 μ g/ml, 49.40 μ g/ml, 1 139 μ g/ml, ID50D3EN-1 respectively 0.031μ g/ml, 25.63 μ g/ml, 1 762μ g/ ml, its developmental toxicity determination in turn were no, weak and strong. Conclusions Based on ITSFn neural development toxicity in vitro models to evaluate the correct judgement of 5 - fluorouraeil, phenytoin and penicillin G developmental toxicity, can be used for in vitro neural developmental toxicity screening and evaluation.

关 键 词：胚胎干细胞 神经发育毒性 有效性 评价模型 

分 类 号：R-332[医药卫生]