摘要
目的探讨在人胚胎干细胞向神经干细胞分化的过程中,叶酸缺乏对分化的影响。方法人胚胎干细胞(human embryonic stem cells,hESCs)H1细胞系培养,并用经拟胚体(embryonic body,EB)法,诱导其向神经干分化。在分化的过程中,分为三组,一组为正常分化对照;一组用甲氨喋呤(Methotrexate,MTX)(可阻断叶酸代谢)处理;一组在MTX处理的同时补充叶酸(Folic acid,FA)。通过免疫荧光方法检测干性标志物Oct-4,Nanog,Sox-2和分化标志分子Tuj-1和Pax6以观察叶酸缺乏对胚胎干细胞分化的影响。结果经EB法成功诱导人胚胎干细胞向神经干细胞分化,可见到神经干分化标志-玫瑰花环,同时免疫荧光结果也表明部分细胞神经分化标志分子Tuj-1和Pax6阳性。MTX组处理后,Tuj-1和Pax6阳性细胞率减少,而MTX+FA分化组,阳性细胞率比MTX组增加,结果P<0.05,有统计学意义。结论 MTX处理导致神经分化细胞减少,补充叶酸可部分拮抗MTX对人胚胎干向神经干分化的抑制。而且人胚胎干细胞向神经干细胞分化的模型是研究叶酸对神经发育影响的合适模型。
Objective： To explore the roles of Folic acid （FA） in neural differentiation in neurogenesis in human embryonic stem cells. Methods： HESCs were induced to differentiate into NSCs with a period of embryonic body （EB） culture under three conditions. Cells in group A underwent EB culture, cells in group B were treated with Methotrexate （MTX） , While the cells in group C treated with MTX and FA.The different markers ware detected by immunofluorescence after hESCS differentiated. Results： In group B, the positive rate of Tuj 1 and Pax6 was lower than that in group A （P〈0.01） . And in group C, the positive rate of Tuj i and Pax6 was higher than that in group A （P〈0.01） . Conclusion： FA depletion decreased cell proliferation and affected neural rosette formation. And the hESCs system is a suitable model for further exploring the mechanism of how FA works in prevention of NTDs.
出处
《中国优生与遗传杂志》
2014年第10期137-139,F0002,共4页
Chinese Journal of Birth Health & Heredity
基金
国家自然科学基金项目号(81300489)
关键词
人胚胎干细胞
神经干细胞
细胞分化
叶酸
甲氨喋呤
Human embryonicstemcells： Neural stemcells
Cell differentiation
Folicacid~ Methotrexate
