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壳聚糖接枝聚乙烯亚胺非病毒载体介导骨形态发生蛋白2基因转染至骨髓间充质干细胞

BMP2 gene delivery to bone mesenchymal stem cell by chitosan-g-PEI nonviral vector.

作者信息

Yue Jianhui, Wu Jun, Liu Di, Zhao Xiaoli, Lu William W

机构信息

Center for Human Tissues and Organs Degeneration, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, 1068 Xueyuan Rd., Shenzhen, 518055 People's Republic of China ; Shenzhen Key Laboratory of Marine Biomedical Materials, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, 1068 Xueyuan Rd., Shenzhen, 518055 People's Republic of China.

Department of Orthopaedic and Traumatology, The University of Hong Kong, 21 Sassoon Rd., Pokfulam, Hong Kong, 999077 People's Republic of China.

出版信息

Nanoscale Res Lett. 2015 Apr 29;10:203. doi: 10.1186/s11671-015-0906-3. eCollection 2015.

Abstract

Nanotechnology has made a significant impact on the development of nanomedicine. Nonviral vectors have been attracting more attention for the advantage of biosafety in gene delivery. Polyethylenimine (PEI)-conjugated chitosan (chitosan-g-PEI) emerged as a promising nonviral vector and has been demonstrated in many tumor cells. However, there is a lack of study focused on the behavior of this vector in stem cells which hold great potential in regenerative medicine. Therefore, in this study, in vitro gene delivering effect of chitosan-g-PEI was investigated in bone marrow stem cells. pIRES2-ZsGreen1-hBMP2 dual expression plasmid containing both the ZsGreen1 GFP reporter gene and the BMP2 functional gene was constructed for monitoring the transgene expression level. Chitosan-g-PEI-mediated gene transfer showed 17.2% of transfection efficiency and more than 80% of cell viability in stem cells. These values were higher than that of PEI. The expression of the delivered BMP2 gene in stem cells enhanced the osteogenic differentiation. These results demonstrated that chitosan-g-PEI is capable of applying in delivering gene to stem cells and providing potential applications in stem cell-based gene therapy.

摘要

纳米技术对纳米医学的发展产生了重大影响。非病毒载体因其在基因递送中的生物安全性优势而受到越来越多的关注。聚乙烯亚胺(PEI)共轭壳聚糖(壳聚糖-g-PEI)作为一种有前景的非病毒载体出现,并已在许多肿瘤细胞中得到证实。然而,缺乏对这种载体在再生医学中具有巨大潜力的干细胞中的行为的研究。因此,在本研究中,研究了壳聚糖-g-PEI在骨髓干细胞中的体外基因递送效果。构建了包含ZsGreen1绿色荧光蛋白报告基因和BMP2功能基因的pIRES2-ZsGreen1-hBMP2双表达质粒,用于监测转基因表达水平。壳聚糖-g-PEI介导的基因转移在干细胞中显示出17.2%的转染效率和超过80%的细胞活力。这些值高于PEI。递送的BMP2基因在干细胞中的表达增强了成骨分化。这些结果表明,壳聚糖-g-PEI能够应用于向干细胞递送基因,并在基于干细胞的基因治疗中提供潜在应用。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8780/4420764/7afa307bcea5/11671_2015_906_Fig1_HTML.jpg

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