THE USE OF EMBRYONIC PRIMATE STEM CELLS IN PARKINSON?S DISEASE MODELS

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。所列机构为中心机构，不一定为研究者机构。如果在体内显示出适当的神经元分化和整合，孤雌生殖干细胞可能成为用于人类患者移植的胎儿组织的有效替代物。在本研究中，我们将研究孤雌生殖灵长类干细胞（Cyno-1，Cibelli et al 2002）在PD动物模型中恢复运动功能的功效。Cyno-1细胞可以维持和扩增很长一段时间，并且使用适当的方案，80%的细胞在体外分化为神经元。这些神经元中的25%表达酪氨酸羟化酶（TH）并释放多巴胺（DA）以响应KCl去极化（Cibelli et al，2002）。在这项研究中，我们将调查这些细胞在体内分化为DA神经元的潜力。将在6-OHDA损伤大鼠模型中进行初步研究，以优化体内条件。我们已经在该模型中使用小鼠干细胞获得了成功的DA分化（Bjokrlund等，2002）。灵长类动物的研究是至关重要的，以评估干细胞衍生的DA神经元的功能功效。在这项研究中，供体和宿主来自相同的物种，这使免疫学问题最小化，但为了鉴定移植的细胞，我们将使用表达GFP的食蟹猴-1细胞。GFP表达没有改变小鼠ES细胞向小鼠DA神经元的分化。此外，我们将移植到雄性宿主中，因此我们将使用Y染色体的原位杂交（FISH）来区分来自Cyno-1（孤雌生殖）的TH神经元与来自宿主的TH-内在纹状体神经元。我们将结合联合收割机行为和功能神经影像学技术，以确定与移植物成熟和整合相关的运动体征的恢复，我们在体内使用正电子发射断层扫描（PET）和突触前和突触后DA标记物的特异性配体进行评估。

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. Parthenogenetic stem cells might become a valid alternative to fetal tissue for transplantation in human patients if appropriate neuronal differentiation and integration are shown in vivo. In this study we will investigate the efficacy of parthenogenetic primate stem cells (Cyno-1, Cibelli et al 2002) to restore motor function in animal models of PD. Cyno-1 cells can be maintained and expanded for long periods of time and using appropriate protocols 80% differentiate into neurons in vitro. Twenty-five percent of these neurons express tyrosine hydroxylase (TH) and release dopamine (DA) in response to KCl depolarization (Cibelli et al, 2002). In this study we will investigate the potential of these cells to differentiate in vivo into DA neurons. Initial studies will be performed in the 6-OHDA-lesioned rat model to optimize the in vivo conditions. We have already obtained successful DA differentiation using mice stem cells in this model (Bjokrlund et al, 2002). Primate studies are critical to evaluate the functional efficacy of stem cell derived DA neurons. In this study donor and host are from the same species, which minimizes immunological problems but in order to identify grafted cells we will use GFP expressing Cyno-1 cells. GFP expression did not alter mouse ES cells differentiation into DA neurons in the mouse. Moreover, we will transplant into male host so we will use in situ hybridization (FISH) for the Y chromosome to differentiate TH neurons derived from Cyno-1 (parthenogenetic) from host derived TH-intrinsic striatal neurons. We will combine behavioral and functional neuroimaging techniques to determine recovery of motor signs in correlation with graft maturation and integration, which we assess in vivo using positron emission tomography (PET) and specific ligands for pre and postsynaptic DA markers.