Kealy Brian, Liew Aaron, McMahon Jill M, Ritter Thomas, O'Doherty Aideen, Hoare Melissa, Greiser Udo, Vaughan Erin E, Maenz Martin, O'Shea Ciara, Barry Frank, O'Brien Timothy
Regenerative Medicine Institute, National Centre for Biomedical Engineering Science & Department of Medicine, National University of Ireland, Galway, Republic of Ireland.
Tissue Eng Part C Methods. 2009 Jun;15(2):223-31. doi: 10.1089/ten.tec.2008.0323.
BACKGROUND/AIMS: The ability of endothelial progenitor cells (EPCs) to home to sites of neoangiogenesis makes them attractive candidates for use in the field of gene therapy. The efficacy of this approach depends on the efficiency of the vector used for transgene delivery.
METHODS/RESULTS: In this study, we have compared the efficiency of adenovirus, five serotypes of AAV2, VSVG-pseudotyped lentivirus, and nonviral plasmid/liposome DNA vectors to deliver the green fluorescence protein reporter gene to human early EPCs to determine efficacy and vector-related cell toxicity. Adenovirus proved most effective with efficiencies of up to 80% with low levels of cell death. Lower levels of expression were seen with other vectors. Electroporation proved unsuitable at the parameters tested. We have also identified at least two distinct subpopulations that exist in the heterogeneous parent EPC culture, one of which is amenable to transduction with adenovirus and one that is not. In addition, adenoviral transduction did not disrupt the ability of the cells to incorporate into endothelial structures in vitro.
We have found adenovirus to be the most efficient of the vector systems tested for gene delivery to EPCs, an effect that is mediated almost entirely by one of two identified subpopulations.
背景/目的：内皮祖细胞（EPCs）归巢至新生血管形成部位的能力使其成为基因治疗领域有吸引力的候选细胞。这种方法的疗效取决于用于转基因递送的载体的效率。
方法/结果：在本研究中，我们比较了腺病毒、五种血清型的AAV2、VSVG假型慢病毒以及非病毒质粒/脂质体DNA载体将绿色荧光蛋白报告基因递送至人早期EPCs的效率，以确定疗效和载体相关的细胞毒性。腺病毒被证明是最有效的，效率高达80%，细胞死亡水平较低。其他载体的表达水平较低。在所测试的参数下，电穿孔被证明不合适。我们还在异质性亲本EPC培养物中鉴定出至少两个不同的亚群，其中一个亚群易于被腺病毒转导，另一个则不然。此外，腺病毒转导并未破坏细胞在体外整合到内皮结构中的能力。
我们发现腺病毒是所测试的用于向EPCs递送基因的载体系统中最有效的，这种效应几乎完全由两个已鉴定的亚群之一介导。
