Cell Grafts for Parkinson's Disease

描述(申请人提供)：神经前体细胞的体外扩增 随后诱导多巴胺能表型的细胞可能提供一个无限的 帕金森氏病(PD)患者移植细胞来源。 然而，控制这些细胞转化为多巴胺的信号 (Da)必须识别神经元。为了做到这一点，单个细胞 从腹侧中脑分离出来的克隆扩增和暴露于 造血细胞因子和神经营养分子。细胞分析 对这种治疗的差异化反应产生了高转化率 某些克隆细胞中酪氨酸羟化酶的百分比(72%至98%) (Th)-阳性表型。在生成的24个克隆中，到 细胞暴露于白介素1(IL-1)的组合中， 白介素11、白血病抑制因子与神经胶质细胞 线源性神经营养因子(GDNF)。阳性克隆表达TH、DA 转运蛋白、NURR-1和在培养中释放DA。暴露于细胞因子中的其他细胞 表达GFAP(星形胶质细胞标志)或MAP-2(神经元标志)的克隆表明 原始神经球也能够产生克隆， 分化为神经胶质细胞和非多巴胺能神经元。初始神经移植 用转化率最高的克隆建立帕金森病大鼠模型的研究 TH表明转化后的祖细胞移植获得了完整的 安非他明诱导的旋转行为的改善并继续 表达TH表型。然而，这些嫁接的成活率 与胚胎腹侧相比，祖细胞减少(26%) 中脑(VM)。这里提出的实验将开发出用于 细胞因子转化的中脑祖细胞的最佳存活。 一旦移植的中脑祖细胞的存活得到优化，直接 在行为测量方面，将与新鲜胚胎VM移植物进行比较， 体内透析，死后DA生化，DA受体，细胞存活和 轴突延伸。最后，这项提案将检验发展议程的效力 胚胎中脑源性克隆祖细胞的转化鸡尾酒 非人灵长类动物的大脑。如果成功，细胞因子转化的中脑 祖细胞有可能取代胚胎组织成为原代细胞 帕金森病移植的细胞来源。

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.