胚胎干细胞实验模型在发育毒性评价中有效性的初步研究

Study on the validation of application model on embryonic stem test

目的依据胚胎干细胞具有向心肌细胞分化的能力,建立体外分化发育毒性评价的模型,并对其判定受试物发育毒性强弱的有效性进行验证。方法利用悬滴、悬浮培养方法,通过不同浓度青霉素G、苯妥英钠和氟尿嘧啶作用,检测胚胎干细胞分化为心肌细胞的能力,结合MTT法获得的受试物细胞毒性结果,判断受试物的发育毒性。结果青霉素G、苯妥英钠和氟尿嘧啶的ID50(D3)分别为1099、47.4和0.023μg/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实验模型能够判定氟尿嘧啶、苯妥英钠和青霉素G发育毒性,结果与ECVAM的结论一致,可用于发育毒性的筛选和评价。

Objective According trend of differentiation cardiomyocyte of embeyonic stem cell,the model of EST has been built and the validation of the model is needed to be tested.It is profit to improve the method of safety evaluation.Methods The embryonic stem cell differentiates into cardiomyocyte in different concentrations of PenicillinG,DPH and 5-FU with hanging and suspending culture conditions.With the results of cytotoxicity,the different embryonictoxicity characteristics of different substants may be detected clearly.Results The ID50（D3） concentration of three embryotoxicants were 1099,47.4 and 0.023μg/ml.PenicillinG,DPH and 5-FU were discriminated as none-embryotoxicity,weak-embryotoxicity and strong-embryotoxicity.Conclusion Test compounds betweem three classes were discriminated correctly.The validation of EST model that established by us is high.