Teramoto S, Ishii T, Matsuse T
Department of Internal Medicine, San-no Hospital, International University of Health and Welfare, Tokyo, Japan.
Hypertens Res. 2001 Jul;24(4):437-43. doi: 10.1291/hypres.24.437.
Factors influencing adeno-associated virus (AAV) - mediated gene transfer to endothelial cells are not fully determined. We tested the variables pertinent to the efficiency of AAV-mediated gene transfer to human vascular endothelial cells (HUVEC) including: (i) kinetics of transduction efficiency of LacZ gene to HUVEC, (ii) the concentration and volume of vector-containing medium, (iii) the period of incubation time of AAV vectors with HUVEC, (iv) the target cell density/proliferation, (v) the duration of transgene expression. There is a dose-response relationship between moi of vectors and transduction efficiency in HUVEC. The higher moi of AAV vectors achieved more than 80% of transduction efficiency in cultured HUVEC. AAV vectors showed incubation time dependent increase in transduction efficiency of LacZ gene to the HUVEC up to 24 h of vector exposure. The foreign gene of AAV vectors preferably transduces the lower density of cells being proliferated. These results indicate that AAV-vector is efficient for gene transfer to HUVEC, and higher moi of vectors or a longer period exposure of vectors to proliferating HUVEC can facilitate efficient tranduction of foreign gene into human vascular endothelial cells in vitro.
影响腺相关病毒（AAV）介导的基因转移至内皮细胞的因素尚未完全明确。我们测试了与AAV介导的基因转移至人血管内皮细胞（HUVEC）效率相关的变量，包括：（i）LacZ基因转导至HUVEC的转导效率动力学；（ii）含载体培养基的浓度和体积；（iii）AAV载体与HUVEC的孵育时间；（iv）靶细胞密度/增殖情况；（v）转基因表达的持续时间。载体的感染复数（moi）与HUVEC中的转导效率之间存在剂量反应关系。较高的AAV载体moi在培养的HUVEC中实现了超过80%的转导效率。在载体暴露长达24小时的时间内，AAV载体显示出LacZ基因转导至HUVEC的转导效率随孵育时间的增加而提高。AAV载体的外源基因更倾向于转导增殖密度较低的细胞。这些结果表明，AAV载体对于基因转移至HUVEC是有效的，并且较高的载体moi或载体对增殖的HUVEC更长时间的暴露可以促进外源基因在体外高效转导至人血管内皮细胞。
