摘要
目的:寻找一种能诱导多种胚胎干细胞向生殖细胞方向分化的化合物,并以常用的诱导物维甲酸(Retinoic Acid,RA)作为阳性参照物。方法:分别培养小鼠胚胎干细胞1B10、D3、R1/E,诱导它们形成拟胚体,让拟胚体贴壁生长,加入特定化合物诱导贴壁细胞分化,诱导72 h后,提取被诱导细胞的RNA,再合成c DNA,最后利用实时荧光定量PCR检测各被诱导细胞中与生殖分化相关的基因的表达水平的变化。结果:发现齐墩果酸(Oleanolic Acid,OA)显著上调了所有被研究的小鼠胚胎干细胞的生殖分化关键基因的表达水平,同时发现阳性参照物RA仅能诱导1B10、R1/E,而不能诱导D3向生殖细胞方向分化。结论:OA能诱导多种胚胎干细胞向生殖细胞方向分化,诱导能力方面具有比常用诱导物维甲酸更广泛的普遍性。
Objective： To find an agent which can induce several strains of embryonic stem cells to differentiate towards germ cells,with the common inducer retinoic acid（ RA） as positive control. Methods： Different strains of mouse embryonic stem cells（ m ESC） 1B10,D3,R1 /E were respectively cultured,and induced to form embryoid bodies（ EBs） which were allowed to attach;different compounds were then added to intervene the attached cells; the intervene continued for 72 h and RNA were extracted and c DNA were synthesized; finally,real-time fluorescence quantitative PCR（ q PCR） assay were used to measure the transcriptional expression patterns of 11 reproductive differentiation related genes. Results： It was found that oleanolic acid（ OA） can induce all 3strains of mouse m ESC to differentiate towards germ cells and the positive control RA can only induce m ESC-1B10 and m ESC-R1 /E to differentiate towards germ cells. RA cannot induce m ESC-D3 to differentiate towards germ cells. Conclusion： OA can induce several strains of embryonic stem cells to differentiate towards germ cells and it has broader universality than the positive control RA.
出处
《世界中医药》
CAS
2015年第2期232-235,242,共5页
World Chinese Medicine
基金
国家重点基础研究发展计划(973计划)"从不孕不育探讨"肾主生殖"理论的研究"的相关研究(编号:2010CB530403)
成都中医药大学实验技术项目(编号:063003)
成都中医药大学校基金(编号:ZRMS201248)
关键词
齐墩果酸
小鼠胚胎干细胞
体外分化诱导物
生殖方向分化
Oleanolic acid
Mouse embryonic stem cells
In vitro differentiation inducer
Differentiation towards germ cells
