INDUCTION OF DOPAMINE NEURONS FROM EMBRYONIC STEM CELLS

该子项目是利用NIH/NCRR资助的中心赠款提供的资源的许多研究子项目之一。子项目和研究者（PI）可能从另一个NIH来源获得主要资金，因此可以在其他CRISP条目中表示。所列机构为中心，不一定是研究者所在机构。探索从猴胚胎干细胞中诱导多巴胺神经元用于帕金森病细胞替代治疗的可能性。 我们建立了一种策略，将多能干细胞导向神经命运。该过程需要在悬浮液中聚集和培养ES细胞，然后在粘附条件下用含有成纤维细胞生长因子2（FGF 2）的混合物分化ES细胞聚集体5-7天。神经前体细胞呈典型的柱状，呈神经管样玫瑰花结状，表达神经上皮标记物巢蛋白和NCAM。通过差异酶反应和差异粘附将这些神经前体细胞与周围的非神经细胞分离至95%纯度。分离的神经前体细胞在体外经FGF 2去除后可分化为神经元和神经胶质细胞。目前正在进行的研究是引导ES衍生的神经前体进一步向多巴胺神经元转化，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。目前正在汇总数据以供发表。

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 monkey embryonic stem (ES) cells for potential cell replacement therapy in 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. The data are being summarized for publication at this time.