DEVELOPMENT OF PRIMATE NEURAL CELLS FOR CNS TRANSPLANTATION

用于中枢神经系统移植的灵长类神经细胞的开发

• 批准号: 3783002
• 负责人: MARY F D NOTTER
• 金额: --
• 依托单位: UNIVERSITY OF ROCHESTER
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3783002
• 关键词: Parkinson's disease; Schwann cells; adrenal medulla; animal tissue; antiparkinson drugs; chromaffin cells; corpus striatum; cytokine; dihydroxyphenylalanine; dopamine; experimental brain lesion; fibroblast growth factor; fibroblasts; histocompatibility; human tissue; in situ hybridization; insulinlike growth factor; laboratory rat; major histocompatibility complex; mixed tissue /cell culture; nervous system transplantation; neurogenesis; neuropharmacology; neurotrophic factors; peripheral nervous system; radioimmunoassay; sciatic nerve; substantia nigra; tissue /cell culture; transfection; vascular endothelium

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.

本提案的总体目标是从体外研究中定义 最佳的灵长类动物细胞营养剂递送系统，可应用于 中枢神经系统（CNS）疾病的临床治疗， 帕金森氏症。 先前的研究表明灵长类动物的肾上腺 可以维持作为候选移植组织的髓质， 当与细胞共培养时， 产生神经营养因子。 因此，在这些拟议的研究中， 将检查来自几个来源的灵长类许旺细胞的特异性 嗜铬细胞生物测定中的神经营养因子，免疫印迹法 和生长因子信使RNA的原位杂交。 永生化的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原种的技术 细胞 将评估这些细胞是否持续产生生长因子 并用作供体细胞，有或没有嗜铬组织， 由Don Gash博士移植到6-羟基多巴胺治疗的CNS中 老鼠，帕金森病的啮齿动物模型。 神经突的基因 酸性成纤维细胞生长因子（AFGF 1）的促进成分将被 插入灵长类动物的细胞， 分化剂 嗜铬细胞，以及其他候选细胞 用afgf 1基因转染的灵长类细胞，将被监测释放 这种营养因子与灵长类嗜铬细胞共培养。 此外，委员会认为， 特异性细胞因子和巨噬细胞衍生因子对 嗜铬细胞的主要组织相容性抗原表达和存活 细胞和雪旺氏细胞将被评估，以开始了解 宿主免疫系统的潜在问题。 永生化细胞 正如所述，将对它们进行审查，因为它们作为股票具有潜力 用于移植的细胞 灵长类嗜铬细胞也会 暴露于不同剂量的左旋多巴，以确定后 移植嗜铬细胞的手术药物治疗。 最终，这些 研究将提供最佳的供体细胞营养剂系统， 可用作退行性CNS疾病的矫正治疗， 帕金森氏症。