Ultrasound-mediated microbubble destruction enhances LMP-1 gene transfection into dendritic cells in vivo.
- Author:
Rui-feng XU
1
;
Min SHI
;
Chang-xuan YOU
;
Cheng-wei LÜ
;
Rong-cheng LUO
;
Wang-jun LIAO
Author Information
- Publication Type:Journal Article
- MeSH: Adaptor Proteins, Signal Transducing; genetics; Cells, Cultured; Contrast Media; administration & dosage; pharmacology; Cytoskeletal Proteins; genetics; Dendritic Cells; drug effects; metabolism; Humans; LIM Domain Proteins; genetics; Microbubbles; Plasmids; Transfection; Ultrasonics
- From: Journal of Southern Medical University 2010;30(10):2327-2332
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the transfection efficiency and the optimal conditions of delivering latent membrane protein-1 (LMP-1) gene to dendritic cells (DCs) by ultrasound exposure combined with contrast agent.
METHODSHuman DCs were cultured in vivo and transfected with the recombinant plasmid pEGFP-C3-LMP1 under varying conditions including ultrasound intensities, exposure time and microbubble contrast agent concentration. The transfection efficiency was assessed by fluorescent microscopy and flow cytometry, and the cell viability by trypan blue exclusion test.
RESULTSAn exposure time of 60 s at MI 1.0 with a microbubble contrast agent concentration of 20% resulted in the optimal effect of delivering the recombinant plasmid pEGFP-C3-LMP1 into the DCs, with a transfection efficiency of (14.37∓2.12)%. Over 90% of the transfected cells were viable after the transfection.
CONCLUSIONMicrobubble contrast agent combined with ultrasound exposure can enhance the delivery of recombinant plasmid pEGFP-C3-LMP1 into the DCs.