Cassel A, Cottler-Fox M, Doren S, Dunbar C E
Clinical Hematology Branch, National Heart, Lung, and Blood Institute, Bethesda, MD 20892.
Exp Hematol. 1993 Apr;21(4):585-91.
Retroviral-mediated gene transfer has been shown to be a feasible method for the introduction of new genes into bone marrow hematopoietic stem cells. We have investigated the application of this technology to primitive CD34-enriched human peripheral blood cells as a potential alternative stem cell source. Bone marrow (BM) and peripheral blood (PB) CD34-enriched cells from normal volunteers and patients with multiple myeloma were exposed to retroviral vectors containing the neomycin-resistance gene and gene transfer efficiency into colony-forming unit colonies (CFU-C) and CD34+ cells was assessed by polymerase chain reaction (PCR). Peripheral blood was a target equally efficient to BM, and PB cells mobilized with chemotherapy and growth factors were also shown to take up retroviral vectors readily. Conditions favoring gene transfer were investigated, and exposure of cells to interleukin-3 (IL-3), interleukin-6 (IL-6), and stem cell factor (SCF) during a 72-hour transduction was found to be most effective. The use of PB stem cells as targets for gene transfer could allow repeated collections and transductions, with obvious advantages over a single BM collection.
逆转录病毒介导的基因转移已被证明是一种将新基因导入骨髓造血干细胞的可行方法。我们研究了将该技术应用于富含原始CD34的人类外周血细胞，作为一种潜在的替代干细胞来源。来自正常志愿者和多发性骨髓瘤患者的骨髓（BM）和外周血（PB）富含CD34的细胞暴露于含有新霉素抗性基因的逆转录病毒载体，通过聚合酶链反应（PCR）评估基因转移到集落形成单位集落（CFU-C）和CD34+细胞中的效率。外周血与骨髓一样是同等有效的靶标，并且用化疗和生长因子动员的PB细胞也显示出易于摄取逆转录病毒载体。研究了有利于基因转移的条件，发现细胞在72小时转导过程中暴露于白细胞介素-3（IL-3）、白细胞介素-6（IL-6）和干细胞因子（SCF）是最有效的。将PB干细胞用作基因转移的靶标可以允许重复采集和转导，与单次骨髓采集相比具有明显优势。
