Cell Grafts for Parkinson's Disease

描述（由申请人提供）：神经祖细胞的体外扩增 细胞，然后诱导多巴胺能表型，可以提供无限的 移植到帕金森病患者体内的细胞来源。 然而，控制这些细胞转化为多巴胺的信号 (DA)神经元必须被识别。为了实现这一目标， 从腹侧中脑分离的克隆扩增并暴露于 造血细胞因子和神经营养分子。分析细胞 响应于这种治疗的分化产生了高的转化率。 在某些克隆中，酪氨酸羟化酶的细胞百分比（72%至98%） （TH）-阳性表型。在所产生的24个克隆中，转化为TH的最佳转化率为100%。 细胞暴露于白细胞介素-1（IL-1）， 白细胞介素-11（IL-11）、白血病抑制因子（LIF）和神经胶质细胞 细胞系源性神经营养因子（GDNF）。阳性克隆表达TH，DA 转运蛋白、Nurr-1和释放的DA。其他细胞暴露于尼古丁 克隆表达GFAP（星形胶质细胞标记物）或MAP-2（神经元标记物），表明 原始的神经球也能够产生克隆， 分化成神经胶质和非多巴胺能神经元。初期神经移植 在PD大鼠模型中使用转化率最高的克隆进行研究， TH表明转化的祖细胞移植物产生了完全的 改善安非他明诱导的旋转行为，并继续 表达TH表型。然而，这些嫁接的成活率 与胚胎腹侧相比， 中脑（VM）。这里提出的实验将制定协议， 最佳的存活率的Wafted甘氨酸转化的中脑祖细胞。 一旦移植的中脑祖细胞的存活被优化， 将在行为测量方面与新鲜胚胎VM移植物进行比较， 体内透析、死后DA生物化学、DA受体、细胞存活和 神经突延伸最后，这项建议亦会考验伤残津贴的成效 对胚胎中脑来源的克隆祖细胞的转化鸡尾酒 非人类灵长类动物的大脑如果成功的话， 祖细胞有可能取代胚胎组织， 用于PD移植的细胞来源。

DESCRIPTION (provided by applicant): In vitro expansion of neural progenitor cells followed by induction of dopaminergic phenotype may provide a limitless source of cells for grafting into patients with Parkinson's disease (PD). However, the signals controlling the conversion of these cells into dopamine (DA) neurons must be identified. In an effort to accomplish this, single cells isolated from ventral mesencephalon were clonally expanded and exposed to hematopoeitic cytokines and neurotrophic molecules. Analysis of cell differentiation in response to this treatment yielded conversion of a high percentage (72 to 98 percent) of cells in some clones to a tyrosine hydroxylase (TH)-positive phenotype. Of the 24 clones generated, the best conversion to TH cells occurred with exposure to a combination of interleukin-1 (IL-1), interleukin- 11 (IL-11), leukemia inhibitory factor (LIF), and glial cell line-derived neurotrophic factor (GDNF). Positive clones expressed TH, the DA transporter, Nurr-1 and released DA in culture. Other cells in cytokine-exposed clones expressed GFAP (astrocyte marker) or MAP-2 (neuron marker) indicating that the original neurospheres were also capable of producing clones that differentiate into glial and nondopaminergic neurons. Initial neural grafting studies m the rat model of PD using a clone with the highest conversion rate to TH indicated that converted progenitor cell grafts produced complete amelioration of amphetamine-induced rotational behavior and continued to express the TH phenotype. However, the survival rate of these grafted progenitor cells was reduced (26 percent) compared to embryonic ventral mesencephalon (VM). The experiments proposed here will develop protocols for optimal survival of Wafted cytokine-converted mesencephalic progenitor cells. Once survival of grafted mesencephalic progenitor cells is optimized, direct comparisons will be made to fresh embryonic VM grafts on measures of behavior, in vivo dialysis, post-mortem DA biochemistry, DA receptors, cell survival and neurite extension. Lastly, this proposal will test the efficacy of the DA conversion cocktail on clonal progenitors derived from embryonic mesencephalon of nonhuman primate brain. If successful, cytokine-converted mesencephalic progenitor cells could potentially replace embryonic tissue as the primary source of cells for grafting in PD.