Baculovirus as a new gene delivery vector for stem cell engineering and bone tissue engineering
Baculovirus as a new gene delivery vector for stem cell engineering and bone tissue engineering
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DOI：
10.1038/sj.gt.3302996
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发表时间：
2007-10-01
期刊：
影响因子：
5.1
通讯作者：
Hu, Y-C
中科院分区：
文献类型：
--
作者：
Chuang, C-K;Sung, L-Y;Hu, Y-C
由于杆状病毒在哺乳动物细胞中的低细胞毒性和不可复制性，杆状病毒已成为一种新的体外和体内基因传递载体，但杆状病毒在人间充质干细胞（hMSCs）遗传修饰和组织工程中的应用尚未见报道。在这项研究中，我们用表达骨形态发生蛋白-2 （BMP-2）的杆状病毒（Bac-CB）对hMSCs进行基因工程改造。造血干细胞在40次感染时的Bac-CB转导触发了造血干细胞向成骨细胞的有效分化。碱性磷酸酶测定、茜素红染色和成骨基因逆转录聚合酶链反应分析证实，初始转导后第6天的超转导增强了BMP-2的表达，进一步加速了体外成骨。苏木精和伊红、免疫组织化学和茜素红染色证实，将超转导细胞植入裸鼠的异位部位，在第2周时细胞有效分化为成骨细胞，并在第6周诱导进行性矿化和部分骨形成。这些数据首次共同证明了杆状病毒在hMSCs工程中的潜力，并暗示了其在骨组织工程中的应用。
Baculovirus has emerged as a novel vector for in vitro and in vivo gene delivery due to its low cytotoxicity and non-replication nature in mammalian cells, but the applications of baculovirus in the genetic modification of human mesenchymal stem cells (hMSCs) and tissue engineering are yet to be reported. In this study, we genetically engineered hMSCs with a baculovirus (Bac-CB) expressing bone morphogenetic protein-2 (BMP-2). Bac-CB transduction of hMSCs at a multiplicity of infection of 40 triggered effective differentiation of hMSCs into osteoblasts. Supertransduction at day 6 after initial transduction enhanced the BMP-2 expression and further accelerated the in vitro osteogenesis, as confirmed by alkaline phosphatase assay, Alizarin red staining and reverse transcription-polymerase chain reaction analysis of osteoblastic genes. Implantation of the supertransduced cells at ectopic sites in the nude mice resulted in efficient cell differentiation into osteoblasts at week 2 and induced progressive mineralization and partial bone formation at week 6, as confirmed by hematoxylin and eosin, immunohistochemical and Alizarin red staining. These data collectively demonstrated, for the first time, the potential of baculovirus in hMSCs engineering and implicated its use in bone tissue engineering.