人孤雌胚胎干细胞向多巴胺能神经元的体外分化  

Differentiation of Dopaminergic Neurons from human parthenogenetic embryonic stem Cells

作　　者：袁丁[1] 卢光琇[1] 林戈[1] 

机构地区：[1]中南大学生殖与干细胞工程研究所(人类干细胞国家工程研究中心),湖南长沙410078

出　　处：《中国现代医学杂志》2010年第3期342-345,共4页China Journal of Modern Medicine

基　　金：国家"863"计划项目(2006AA02A102)

摘　　要：目的研究人孤雌胚胎干细胞向神经上皮祖细胞和神经元诱导分化的能力。方法人孤雌胚胎干细胞在人饲养层细胞上诱导后机械消化传代,在多巴胺能神经元诱导培养基中悬浮后贴壁分化,Pax6、Nestin和PSA-NCAM抗原染色检测神经上皮祖细胞的形成,Tuj1和TH抗原染色检测多巴胺能神经元的形成。结果人孤雌胚胎干细胞诱导可形成的神经上皮祖细胞中,具有典型的rosette结构,其Pax6和Nestin阳性细胞比例分别为(84.7±5.3)%和(92.0±4.7)%。该神经上皮祖细胞可形成PSA-NCAM染色阳性的神经元前体和Tuj1染色阳性的神经元,Tuj1阳性细胞中(33.3±4.3)%为酪氨酸羟化酶染色阳性细胞。结论人孤雌胚胎干细胞体外诱导分化可形成高纯度的神经上皮祖细胞,并可进一步分化为多巴胺能神经元。【Objective】To study the differentiating ability of human parthenogenetic embryonic stem cells to neuroepithelial progenitor cells and neurons.【Methods】Human parthenogenetic embryonic stem cells cultured on human feeder cells were induced and mechanically passaged,then floated and replated in dopaminergic neuronal inducing medium for further differentiation.Pax6,Nestin and PSA-NCAM were used as antigen markers of neuroep-ithelial progenitor cell genesis,and Tuj1 and TH of dopaminergic neuron genesis.【Results】Human parthenogenetic embryonic stem cells could differentiated into neuroepithelial progenitor cells with typical ＇rosette＇ structure,the per-centage of Pax6 and Nestin positive cells of which were（84.7±5.3） % and（92.0±4.7） % respectively.Derived neuroepithelial progenitor cells could differentiate into PSA-NCAM positive neuronal precursors and Tuj1 positive neurons,and（33.3±4.3）% Tuj1 positive cells were tyrosine hydroxylase positive cells.【Conclusions】Human parthenogenetic embryonic stem cells could be differentiated in vitro into highly purified neuroepithelial progenitor cells,and further into dopaminergic neurons.

关 键 词：孤雌胚胎干细胞 神经诱导 多巴胺能神经元 

分 类 号：Q2-33[生物学—细胞生物学] Q21