Gene transfer into rat mesenchymal stem cells: A comparative study of viral and nonviral vectors
Gene transfer into rat mesenchymal stem cells: A comparative study of viral and nonviral vectors
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DOI：
10.1089/scd.2006.15.87
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发表时间：
2006-02-01
影响因子：
4
通讯作者：
O'Brien, T
中科院分区：
文献类型：
--
作者：
McMahon, JM;Conroy, S;O'Brien, T
间充质干细胞（MSC）已被提议用于组合基因和细胞治疗方案，以治疗疾病和促进修复。这种治疗方法的功效取决于确定哪些载体以最小的细胞死亡提供最大的转基因表达。该研究在大鼠骨髓间充质干细胞上进行，并在相同的干细胞制剂上测试了一系列载体。比较了腺病毒、腺相关病毒（AAV；血清型 1、2、4、5 和 6）、慢病毒和非病毒载体。慢病毒被证明是最有效的，转导效率高达 95%，同时细胞毒性水平较低。腺病毒也被证明是有效的，但随着病毒滴度的增加，细胞死亡显着增加。与同时测试的兔 MSC 相比，大鼠 MSC 对所有 AAV 血清型的转导仍然无效。质粒DNA的脂转染产生中等转染水平，但也伴随着细胞死亡。电穿孔基因转移在测试参数下被证明是无效的，并导致大量细胞死亡。使用慢病毒载体进行高水平和中等水平的细胞转导并不影响细胞分化成脂肪途径的能力。
Mesenchymal stem cells (MSCs) have been proposed for use in combinatorial gene and cell therapy protocols for the treatment of disease and promotion of repair. The efficacy of such a therapeutic approach depends on determination of which vectors give maximal transgene expression with minimal cell death. The study was carried out on bone-marrow derived rat MSCs, and a range of vectors was tested on the same stem cell preparation. Adenovirus, adeno-associated virus (AAV; serotypes 1, 2, 4, 5, and 6), lentivirus, and nonviral vectors were compared. Lentivirus proved to be most effective with transduction efficiencies of up to 95%, concurrent with low levels of cell toxicity. Adenovirus also proved effective, but a significant increase in cell death was seen with increasing viral titer. Rat MSCs remained refractory to transduction by all AAV serotypes, in contrast to rabbit MSCs tested at the same time. Lipofection of plasmid DNA gave moderate transfection levels but was also accompanied by cell death. Electroporative gene transfer proved ineffective at the parameters tested and resulted in high cell death. High and moderate levels of cell transduction using lentivirus vectors did not affect the ability of the cells to differentiate down the adipogenic pathway.