人脐血干细胞定向分化为多巴胺能神经元的实验研究  

EXPERIMENTAL STUDY ON DIRECTIONAL DIFFERENTIATION OF HUMAN UMBILICAL CORD BLOOD(HUCB) STEM CELLS INTO DOPAMINERGIC NEURONS

作　　者：李荣平[1] 季凤清[1] 孙海梅[1] 王屹[2] 曾晓蓓[1] 赵春礼[2] 杨慧[2] 李雪梅[3] 贾晓芳[3] 

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 [3]首都医科大学附属天坛医院妇产科,北京100069

出　　处：《解剖学报》2007年第2期148-152,共5页Acta Anatomica Sinica

基　　金：北京市优秀人才培养基金(20042D0501805);北京市教委科技发展计划面上项目(KM200610025005)资助课题

摘　　要：目的探讨人脐血干细胞定向分化为多巴胺(DA)能神经元的最佳诱导条件。方法体外分离、原代培养人脐血干细胞,流式细胞仪检测其表面标志,分别以EGF+bFGF、ATRA、ATRA+EGF+bFGF、纹状体条件培养液、纹状体星形胶质细胞条件培养液对其进行诱导,倒置相差显微镜观察细胞形态的变化。应用免疫荧光染色技术检测DA能神经元标志物酪氨酸羟化酶(TH)的表达。结果比较各组对人脐血干细胞定向分化为DA能神经元的诱导作用,纹状体星形胶质细胞条件培养液>纹状体条件培养液>ATRA+EGF+bFGF组>ATRA组>EGF+bFGF组>对照组。结论纹状体组织对人脐血干细胞定向分化为DA能神经元的诱导作用优于其他各组,并且此作用可能主要源于纹状体的星形胶质细胞。Objective To investigate and compare some conditions which could induce human umbilical cord blood （HUCB） stem cells to differentiate into dopaminergic neurons, and probe the optimal condition for the directional differentiation of HUCB stem cells. Methods HUCB stem cells were isolated and cultured in vitro, and the cell surface markers were detected by flow cytometry, and when the cells were co-cultured with EGF ＋ bFGF, ATRA, ATRA ＋ EGF ＋ bFGF, striatal conditioned medium（CM ）, striatal astrocytes conditioned medium（ CM ） respectively, the morphological character was observed by inverse phase-contrast microscopy. The expression of tyrosine hydroxylase （TH） was detected by immunofluorescence staining method. Results The comparison of the results of HUCB stem cells which directly differentiated into dopaminergic neuron showed that the differentiation potential of striatal astrocytes CM group 〉 striatal CM group 〉 ATRA ＋ EGF ＋ bFGF group 〉 ATRA group 〉 EGF ＋ bFGF group 〉 control group. Conclusion Compared with other conditions, the striatum has a more distinct inductive effect on HUCB stem cells to differentiate into dopaminergic neurons, and the inductive effect mainly comes from the astrocytes in the striatum.

关 键 词：干细胞 诱导分化 多巴胺能神经元 纹状体 星形胶质细胞 免疫细胞化学 人脐带血 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学]