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

为了开发一种临床前基因治疗模型，使用 CD34造血干细胞，我们比较了不同的方法 CD34干细胞在恒河猴体内的获取和移植。我们 研究重组人粒细胞集落刺激因子和干细胞因子动员CD34的能力 细胞进入外周循环。我们通过以下方式收集了这些 血浆障碍症。我们还从骨髓中收集了CD34细胞 菲科尔分离。后一种细胞被逆转录病毒转导。 含有报告基因的载体，并回输给供体 猴子。接受回输的猴子全身接受200cGy射线照射 照射，以净化骨髓，提高机会 刻好了。第一批获得转导细胞的猴子被重新植入 仅表现出轻微的转基因表达。

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.