DEVELOPMENT OF PRECLINICAL MODEL FOR GENE THERAPY USING CD34+ STEM CELLS

使用 CD34 干细胞进行基因治疗的临床前模型的开发

• 批准号: 6247290
• 负责人: GARY B BASKIN
• 金额: $ 6.02万
• 依托单位: TULANE UNIVERSITY OF LOUISIANA
• 依托单位国家: 美国
• 财政年份: 1997
• 资助国家: 美国
• 起止时间: 1997-05-09 至 1998-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6247290
• 关键词: Mammalia; Primates; biotechnology; blood; gene therapy; genetics; model design /development; technology /technique development; virus

In order to develop a preclinical model of gene therapy using CD34+ hematopoietic stem cells, we compared various methods of obtaining and engrafting CD34+ stem cells in rhesus monkeys. We investigated rhG-CSF and rhSCF for their ability to mobilize CD34+ cells into the peripheral circulation. We collected these by plasmapharesis. We also collected CD34+ cells from bone marrow by ficoll separation. The latter cells were transduced with a retroviral vector containing a reporter gene, and were reinfused into the donor monkeys. Monkeys to be reinfused receive 200 cGy total body irradiation in order to purge the marrow and improve the chances of engraftemnt. The first monkeys to have transduced cells reinoculated have shown only slight transgene expression.

为了开发基因治疗的临床前模型 CD34+造血干细胞，我们比较了各种方法 在恒河猴中获得并移植 CD34+ 干细胞。 我们 研究了 rhG-CSF 和 rhSCF 动员 CD34+ 的能力 细胞进入外周循环。 我们收集了这些 血浆去除术。 我们还通过以下方法从骨髓中收集了 CD34+ 细胞： 聚蔗糖分离。 后者的细胞被逆转录病毒转导 含有报告基因的载体，并被回输到供体中 猴子。 接受回输的猴子全身接受 200 cGy 辐射以净化骨髓并提高机会 植入。 第一批将转导细胞重新接种的猴子 仅表现出轻微的转基因表达。