论文部分内容阅读
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: To establish an evaluation model of neurodevelopmental toxicity of embryonic stem cells based on insulin-transferrin-selenium-fibronectin (ITSFn) culture in vitro and validate its neurotoxicity to evaluate the neurodevelopmental toxicity. METHODS: Embryonic stem cells were suspended in suspension and cultured in vitro. Insulin, transferrin, selenium and fibronectin were added to induce neuronal differentiation. Neurotoxicity models were established by cytotoxic MTT in vitro. The effects of different concentrations of penicillin G, phenytoin and Fluorouracil under the conditions of its ability to differentiate embryonic stem cells to determine the neurotoxicity of test substances. Results The ID50D3 nestin of penicillin G, phenytoin and fluorouracil were 0.017μg / ml, 49.40μg / ml, 1 139μg / ml and ID50D3EN-1 respectively 0.031μg / ml, 25.63μg / ml and 1 762μg / ml. Judgments were followed by no, weak and strong. Conclusion The in vitro neurodevelopmental toxicity evaluation model based on ITSFn induction method can correctly determine the developmental toxicity of fluorouracil, phenytoin sodium and penicillin G, and can be used to screen and evaluate the neurotoxicity in vitro.