羊膜对人脐血干细胞分化为多巴胺能神经元的作用  

THE FUNCTION OF AMNION IN TNE DIFFERENTIATION OF HUMAN UMBILICAL CORD BLOOD STEM CELLS INTO DOPAMINERGIC NEURONS

作　　者：李荣平[1] 季凤清[1] 孙海梅[1] 王丹妮[1] 曾小蓓[1] 赵春礼[2] 杨慧[2] 

机构地区：[1]首都医科大学组织学胚胎学教研室 [2]首都医科大学神经科学研究所,北京100069

出　　处：《解剖学报》2008年第6期790-794,共5页Acta Anatomica Sinica

基　　金：国家重点基础研究发展计划(973)(2006CB500706);北京市教委科技发展计划(KM200610025005)资助项目

摘　　要：目的探讨羊膜上皮细胞对人脐血干细胞定向分化为多巴胺能神经元的作用。方法体外分离、原代培养人羊膜上皮细胞,通过高速离心制备成条件培养液(CM)对P1代脐血干细胞进行诱导,倒置相差显微镜观察细胞形态的变化,应用免疫荧光染色和免疫印迹法检测其酪氨酸羟化酶(TH)及多巴胺转运体(DAT)蛋白的表达情况。结果P1代脐血干细胞经羊膜上皮细胞条件培养液诱导后,TH及DAT阳性细胞率明显高于对照组,与对照组比较有显著性差异(P<0.01),免疫印迹分析结果与免疫荧光染色结果一致。结论羊膜上皮细胞能诱导人脐血间充质干细胞定向分化为多巴胺能神经元。Objective To investigate the function of amnion endothelial cell in the differentiation of human umbilical cord blood stem cells into dopaminergic neurons. Methods Primary human amnion endothelial cells were separated and cultured in vitro ; the conditioned medium（CM） was prepared through high speed centrifugation. The cord blood mesenchymal stem cells of Pl passage were induced by the conditioned medium, and the mophology of cells was observed under the inverted phase contrast microscope. The expression of tyrosine hydroxylase（TH）and dopamine transportor （DAT） of the induced cord blood mesenchymal stem cells were detected by immunocytochemistry staining method and immunoblotting（Western blotting）, Results The masculine rate of TH and DAT of the cord blood mesenchymal stem cells of Pl passage which were induced by amnion endothelial cells conditioned medium was higher than that of the control group, with a significant difference （P 〈 0.01 ）. The results of Western blotting were in agreemctn with the immunocytochemistry staining. Conclusion Amnion endothelial cells can induce human umbilical cord blood stem cells into dopaminergic neurons.

关 键 词：干细胞 诱导分化 多巴胺能神经元 羊膜上皮细胞 免疫组织化学 人脐带血 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学]