Manufacturing human Neural Stem & Progenitor Cells under reduced oxygen.

制造人类神经干

• 批准号: 7107752
• 负责人: SMITA I SAVANT-BHONSALE
• 金额: $ 16.53万
• 依托单位: THERADIGM, INC.
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-05 至 2007-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7107752
• 关键词: brain; conditioning; embryo /fetus; human; oxygen; spinal cord; stem cells

DESCRIPTION (provided by applicant): Theradigm has a goal to develop and deliver cell based therapy for neurological disease or injury. In recent years tremendous advances are made in the field of cell therapy to repair injured tissue. But the cell therapy field still faces the challenges of consistently expanding the therapeutic cells in large enough quantities without immortalizing them for their use in human clinical trials. We are proposing to manufacture human fetal brain and spinal cord derived neural stem cells (NSCs) under physiological oxygen levels (3%). Based on our preliminary data and other research reports we hypothesize that expansion of NSCs under lower than atmospheric oxygen (20%) will increase their expansion rate while maintaining their multipotency. We will first identify the most optimum growth conditions to expand these cells in vitro. We will then characterize these cells for their stem cell properties by looking for expression of stem cell specific markers by Flow cytometry and quantitative PCR. Their multipotential by differentiating them and measuring expression of brain lineage specific genes by quantitative PCR. We will define their phenotype by gene profile on microarrays and expression of cell surface markers by Flow cytometry. By high-resolution array- based copy number analysis, HLA typing, SIR typing and karyotyping we will determine genomic stability of NSCs grown under reduced oxygen. After defining their identity (molecular, phenotypic, immunologic, genotypic) we will use the optimum growth conditions to manufacture and bank these cells. These well characterized cells will be used in Phase II study to determine engraftment and efficacy of these cells in animal spinal cord injury models. These studies will help us with our long-term goal to develop cell therapy to improve morbidity and health care cost for patients suffering from spinal cord injury. If we can manufacture NSCs in large enough quantities, we can also test efficacy of them in other neurological disorders and injuries like stroke and traumatic brain injury. We will also be able to apply this cell manufacturing process to other cell types like bone marrow and adipose tissue derived stem cells.

描述（由申请人提供）：Theradigm的目标是开发和提供用于神经系统疾病或损伤的细胞疗法。近年来，在细胞治疗领域取得了巨大的进展，以修复受损组织。但是细胞治疗领域仍然面临着不断扩大治疗细胞的挑战，这些治疗细胞的数量足够大，但不能使它们永生化，以用于人体临床试验。我们建议在生理氧水平（3%）下制造人胎脑和脊髓来源的神经干细胞（NSCs）。根据我们的初步数据和其他研究报告，我们假设在低于大气氧气（20%）的条件下NSC的扩增将增加其扩增速率，同时保持其多能性。我们将首先确定在体外扩增这些细胞的最佳生长条件。然后，我们将通过流式细胞术和定量PCR寻找干细胞特异性标志物的表达来表征这些细胞的干细胞特性。通过区分它们并通过定量PCR测量脑谱系特异性基因的表达来检测它们的多潜能。我们将通过微阵列上的基因谱和流式细胞术检测细胞表面标志物的表达来确定其表型。通过基于高分辨率阵列的拷贝数分析、HLA分型、SIR分型和核型分析，我们将确定在低氧下生长的NSC的基因组稳定性。在确定其身份（分子、表型、免疫学、基因型）后，我们将使用最佳生长条件生产和储存这些细胞。这些充分表征的细胞将用于II期研究，以确定这些细胞在动物脊髓损伤模型中的植入和功效。这些研究将帮助我们实现开发细胞疗法的长期目标，以改善脊髓损伤患者的发病率和医疗保健成本。如果我们能够大量制造NSC，我们还可以测试它们在其他神经系统疾病和损伤（如中风和创伤性脑损伤）中的疗效。我们还将能够将这种细胞制造过程应用于其他细胞类型，如骨髓和脂肪组织来源的干细胞。