Genetically engineered angiogenic cell sheets using magnetic force-based gene delivery and tissue fabrication techniques
Genetically engineered angiogenic cell sheets using magnetic force-based gene delivery and tissue fabrication techniques
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DOI：
10.1016/j.biomaterials.2009.11.017
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发表时间：
2010-02-01
期刊：
影响因子：
14
通讯作者：
Kamihira, Masamichi
中科院分区：
文献类型：
--
作者：
Akiyama, Hirokazu;Ito, Akira;Kamihira, Masamichi
组织工程的一个主要限制是在移植组织中血管的形成不足，导致细胞密度和移植物大小的降低。我们在这里报告了血管生成细胞片的制造，使用两种磁力技术的组合，其中使用磁铁矿阳离子脂质体（mcl），磁感染和磁性细胞积累。用mcl标记编码血管内皮生长因子（VEGF）表达盒的逆转录病毒载体，以磁性吸引颗粒到小鼠成肌细胞C2C12单层上，用于基因传递。与传统方法相比，mcl介导的感染使转导效率提高了6.7倍。在组织结构的制造过程中，mcl标记的细胞在磁场的存在下积累，以促进多层细胞片的自发形成。VEGF基因工程C2C12 （C2C12/VEGF）细胞片，采用磁力技术构建，皮下移植到裸鼠体内。组织学分析显示，在第14天，C2C12/VEGF细胞片产生了较厚的组织，细胞密度高，并促进了血管化。这表明这里描述的方法代表了组织工程中的一种强有力的策略。2009爱思唯尔有限公司版权所有。
A major limitation in tissue engineering is the insufficient formation of blood vessels in implanted tissues, resulting in reduced cell density and graft size. We report here the fabrication of angiogenic cell sheets using a combination of two magnetic force-based techniques which use magnetite cationic liposomes (MCLs), magnetofection and magnetic cell accumulation. A retroviral vector encoding an expression cassette of vascular endothelial growth factor (VEGF) was labeled with MCLs, to magnetically attract the particles onto a monolayer of mouse myoblast C2C12 cells, for gene delivery. MCL-mediated infection increased transduction efficiency by 6.7-fold compared with the conventional method. During the fabrication of the tissue constructs, MCL-labeled cells were accumulated in the presence of a magnetic field to promote the spontaneous formation of a multilayered cell sheet. VEGF gene-engineered C2C12 (C2C12/VEGF) cell sheets, constructed using both magnetic force-based techniques, were subcutaneously transplanted into nude mice. Histological analyses revealed that on day 14 the C2C12/VEGF cell sheet grafts had produced thick tissues, with a high-cell density, and promoted vascularization. This suggests that the method described here represents a powerful strategy in tissue engineering. (C) 2009 Elsevier Ltd. All rights reserved.