应用胚胎干细胞试验模型对碱性成纤维细胞生长因子发育毒性的初步评价

摘要

目的 利用小鼠胚胎干细胞试验(EST)模型,初步评价bFGF的发育毒性.方法 体外培养小鼠胚胎干细胞(ESC),通过形态学观察、碱性磷酸酶染色和RT-PCR方法进行ESC未分化鉴定;MTT法检测不同浓度bFGF对ESC和3T3细胞的毒性,RT-PCR半定量分析法检测bFGF对未分化基因Sox-2表达的影响.结果 bFGF对ESC和3T3的半数抑制浓度和ESC的半数抑制分化浓度分别为IC50 ESC=15.2 mg·L-1,IC50 3T3=24.2 mg·L-1,ID50 ESC=1.7 mg·L-1.结论 bFGF发育毒性的判定为弱胚胎毒性.%Aim To research the developmental toxicity of basic Fibroblast Growth Factor ( bFGF ) by theembryonic stem cell test. Methods Mouse embryonic stem cells were cultured and their undifferentiationswere identified by the methods of morphology, alkaline phosphatase staining and RT-PCR. The cytotoxic effect of bFGF was detected by MTT assay. The effect of bF-GF on Sox-2 gene expression was determined by semi-quantitative RT-PCR. Result The IC50 of bFGF on3T3 was 24. 2 mg · L-1, while IC50 and ID50 on ESCwere 15. 2 mg · L -1 and 1. 7 mg · L -1 respectively. Conclusion The developmental toxicity of bFGF is e-valuated as weak embryotoxicity.