INDUCTION OF DOPAMINE NEURONS FROM EMBRYONIC STEM CELLS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 探索从无限来源的人类和非人类灵长类胚胎干细胞中产生多巴胺神经元的可能性，以研究治疗帕金森病的潜在细胞替代疗法。 我们建立了一种策略，将多能干细胞引向神经的命运。这一过程需要在悬浮液中聚集和培养ES细胞，然后在含有成纤维细胞生长因子2(FGF2)的鸡尾酒中贴壁条件下分化ES细胞聚集体5-7天。神经前体细胞呈典型的柱状形态，排列成神经管状玫瑰花环，表达神经上皮细胞标志物Nestin和NCAM。通过差异性酶反应和差异性黏附，将神经前体细胞从周围的非神经细胞中分离出来，纯度达到95%。体外分离的神经前体细胞在FGF2去除后分化为神经元和神经胶质细胞。正在进行的研究是将ES来源的神经前体进一步引导到多巴胺神经元，并在帕金森病大鼠模型中检测多巴胺神经元的功能。这项研究使用了WNPRC干细胞资源和联邦批准的HES细胞系。 资金说明：私人资金。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. To explore the possibility of generating dopamine neurons from an unlimited source of human and nonhuman primate embryonic stem (ES) cells to study potential cell replacement therapies for Parkinson's disease. We established a strategy that directs the pluripotent stem cells to a neural fate. This process entails aggregation and culture of ES cells in suspension, followed by differentiation of the ES cell aggregates in an adherent condition with a cocktail containing fibroblast growth factor 2 (FGF2) for 5-7 days. Neural precursor cells displayed a characteristic columnar morphology, organized into neural tube-like rosette formations, and expressed neuroepithelial marker, nestin and NCAM. These neural precursor cells were separated from surrounding non-neural cells to 95% purity by differential enzymatic response and differential adhesion. The isolated neural precursor cells differentiated into neurons and glial cells after withdraw of FGF2 in vitro. Ongoing study is to guide the ES-derived neural precursors further to dopamine neurons and examine the function of the dopamine neurons in a rat model of Parkinson's disease. This Research used WNPRC Stem Cell Resources and federally approved hES cell lines. FUNDING NOTE: Private funding.