Ultrasound-targeted microbubble destruction enhances AAV mediated gene transfection: human RPE cells in vitro and the rat retina in vivo
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- Department of Ultrasound, Department of Ultrasound, Shanghai Jiaotong University Affiliated First People's Hospital
- Department of Ultrasound, Department of Ultrasound, Shanghai Jiaotong University Affiliated First People’s Hospital
- Experimental Research Center of Shanghai Jiaotong University Affiliated First People's Hospital
- Department of Ultrasound, Shanghai Jiaotong University Affiliated First People’s Hospital
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- Date:
- Received 24 February 2009 07:29 UTC; Posted 26 February 2009
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- Biotechnology
- Abstract:
The present study was performed to investigate the efficacy and safety of Ultrasound-targeted microbubble destruction (UTMD) mediated rAAV2-EGFP to cultured human retinal pigment epithelium (RPE) cells in vitro and the rat retina in vivo. In vitro study, cultured human RPE cells were exposed to US under different conditions with or without microbubbles. Furthermore, the effect of UTMD to rAAV2-EGFP itself and the cells were evaluated. In vivo study, gene transfer was examined by injecting rAAV2-EGFP into the subretinal space of the rats with or without microbubbles and then exposed to US. We investigated EGFP expression in vivo via stereomicroscopy and performed quantitative analysis by Axiovision 3.1 software. HE staining and frozen sections were used to observe tissue damage and location of EGFP gene expression. In vitro study, the transfection efficiency of rAAV2-EGFP increased 74.85% under the optimal UTMD conditions. Furthermore, there was almost no cytotoxicity to the cells and rAAV2-EGFP itself. In vivo study, UTMD could be used safely to enhance and accelerate transgene expression of the retina. Fluorescence expression was mainly located in the layer of retina. UTMD is a promising method for gene delivery to the retina.
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- This document is licensed to the public under the Creative Commons Attribution 3.0 License
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Li, Hongli, Zheng, Xiaozhi, Wang, Huiping, Li, Fan, Wu, Ying, and Du, Lianfang. Ultrasound-targeted microbubble destruction enhances AAV mediated gene transfection: human RPE cells in vitro and the rat retina in vivo. Available from Nature Precedings <http://hdl.handle.net/10101/npre.2009.2898.1> (2009)
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