INDUCTION OF DOPAMINE NEURONS FROM EMBRYONIC STEM CELLS

本子项目是利用由NIH/NCRR资助的中心赠款提供的资源的众多研究子项目之一。子项目和研究者（PI）可能已经从另一个NIH来源获得了主要资金，因此可以在其他CRISP条目中表示。列出的机构是中心的，不一定是研究者的机构。探索从无限来源的猴胚胎干（ES）细胞中产生多巴胺神经元的可能性，用于帕金森病的潜在细胞替代治疗。我们建立了一种策略来引导多能干细胞走向神经的命运。这个过程需要在悬浮中聚集和培养胚胎干细胞，然后在含有成纤维细胞生长因子2 （FGF2）的混合物的贴壁条件下分化胚胎干细胞聚集体5-7天。神经前体细胞呈柱状，组织成神经管样玫瑰花状，表达神经上皮标志物、巢蛋白和NCAM。这些神经前体细胞通过不同的酶反应和不同的粘附从周围的非神经细胞中分离出来，纯度达到95%。体外提取FGF2后，分离的神经前体细胞分化为神经元和胶质细胞。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，并检查多巴胺神经元在帕金森病大鼠模型中的功能。本研究使用WNPRC干细胞资源。目前正在对这些数据进行总结以供出版。

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.