DEVELOPMENT OF PRIMATE NEURAL CELLS FOR CNS TRANSPLANTATION

这项提案的总体目标是从体外研究中定义 最优灵长类细胞营养剂给药系统 中枢神经系统(CNS)疾病的临床治疗 帕金森氏症。此前的研究表明，灵长类肾上腺 髓质是一种候选的移植组织，可以维持和 在体外和体内与细胞共同培养时，表型发生了变化 产生神经营养因子。因此，在这些拟议的研究中， 来自几个来源的灵长类雪旺细胞将被检查出特定的 免疫印迹法检测嗜铬细胞中的神经营养因子 用原位杂交技术检测生长因子(S)信使核糖核酸。 永生灵长类许旺细胞将通过基因转移产生 获得分泌生长因子的组织培养基库的技术 细胞。这些细胞将被评估为持续产生生长因子。 并用作供体细胞，有或没有嗜铬组织，用于 唐·加什博士经6-羟基多巴胺治疗的中枢神经系统移植 大鼠，帕金森氏症的啮齿动物模型。神经突起的基因 酸性成纤维细胞生长因子(Afgf1)的促进成分将 插入灵长类细胞以提供自分泌来源 差异剂。嗜铬细胞，以及其他候选细胞 将监测转入afgf1基因的灵长类细胞释放 这种营养因子与灵长类嗜铬细胞共培养。此外， 特异性细胞因子和巨噬细胞衍生因子对血管内皮细胞生长的影响 主要组织相容性抗原表达与嗜铬细胞存活 将对细胞和雪旺细胞进行评估，以开始了解 宿主免疫系统的潜在问题。永生化细胞在 正如所述，当它们提供作为股票的潜力时，将对其进行检查 用于移植的细胞。灵长类嗜铬细胞也将 暴露于不同剂量的左旋多巴，以建立后- 嗜铬细胞移植的外科药物治疗。最终，这些 研究将提供最佳的供体细胞营养剂系统，可以 作为矫正疗法应用于退行性中枢神经系统疾病，如 帕金森氏症。

The overall goal of this proposal is to define from in vitro studies the optimal primate cell-trophic agent delivery system that can be applied to clinical treatment of central nervous system (CNS) disorders such as Parkinson's disease. Previous studies have shown that primate adrenal medulla, a candidate transplant tissue, can be maintained and phenotypically altered in vitro and in vivo when cocultured with cells that produced neurotrophic factors. Therefore, in these proposed studies, primate Schwann cells from several sources will be examined for specific neuronotrophic factors in a bioassay with chromaffin cells, by immunoblots and by in situ hybridization for growth factor(s) messenger RNA. Immortalized primate Schwann cells will be generated by gene transfer techniques to obtain a tissue culture stock of growth factor secreting cells. These cells will be assessed for continued growth factor production and used as donor cells, with or without chromaffin tissue, for transplantation by Dr. Don Gash into the CNS of 6-hydroxydopamine-treated rats, the rodent model of Parkinson's disease. The gene for the neurite promoting component of acidic fibroblast growth factor (afgf1) will be inserted into primate cells to provide an autocrine source of differentiating agent. Chromaffin cells, as well as other candidate primate cells transfected with afgf1 gene, will be monitored for release of this trophic factor by coculture with primate chromaffin cells. Moreover, the effect of specific cytokines and macrophage derived factors on the major histocompatibility antigen expression and survival of chromaffin cells and Schwann cells will be assessed to begin to understand the potential problems of the host immune system. Immortalized cells in particular will be examined as they offer potential, as stated, as a stock of cells for transplantation. Primate chromaffin cells will also be exposed to varying doses of levodopa to establish the effect of post- surgery drug therapy on transplanted chromaffin cells. Ultimately, these studies will provide the optimal donor cell trophic agent system that can be applied as corrective therapy in degenerative CNS disorders such as Parkinson's disease.