tailieunhanh - Sonoporation-mediated gene transfer into adult rat dorsal root ganglion cells

Gene transfer into many cell types has been successfully used to develop alternative and adjunct approaches to conventional medical treatment. However, effective transfection of postmitotic neurons remains a challenge. The aim of this study was to develop a method for gene transfer into rat primary dorsal root ganglion neurons using sonoporation. Methods: Dissociated cells from adult rat dorsal root ganglion (DRG) cells were sonicated for 1-8 s at W to determine the optimal ultrasound duration and power for gene transfection and cell survival. Transfection efficiency was compared between sonoporation, liposome and. | Lin et al. Journal of Biomedical Science 2010 17 44 http content 17 1 44 a NSC Tha cost of publication In Journal of Blomodlcal Science Is bome by tlM National Science Council Taiwan JOURNAL OF BIOMEDICAL SCIENCE RESEARCH Open Access SorJoporation-mediated gene transfer into adult rat dorsal root ganglion cells Chung-Ren Lin 1 3 Kuan-Hung Chen1 Chien-Hui Yang1 Jiin-Tsuey Cheng4 Shyr-Ming Sheen-Chen2 Chih-Hsien Wu1 4 Wei-Dih Sy1 and Yi-Shen Chen1 Abstract Background Gene transfer into many cell types has been successfully used to develop alternative and adjunct approaches to conventional medical treatment. However effective transfection of postmitotic neurons remains a challenge. The aim of this study was to develop a method for gene transfer into rat primary dorsal root ganglion neurons using sonoporation. Methods Dissociated cells from adult rat dorsal root ganglion DRG cells were sonicated for 1-8 s at W to determine the optimal ultrasound duration and power for gene transfection and cell survival. Transfection efficiency was compared between sonoporation liposome and lentiviral vector gene transfer techniques. Results The optimum ultrasound intensity was 5 W for 2 s and yielded an efficiency of gene transfection of 31 and a survival rate of 35 . Conclusions Sonoporation can be optimized to minimize cell death and yield a high percentage of transfected neurons and that this technique can be easily applied to primary cultures of rat dorsal root ganglion neurons. Background Methods for altering gene expression are widely used to elucidate molecular mechanisms involved in cellular physiopathology. Gene modification also has potential as a therapeutic modality for treating many diseases. Many gene transfer methods have been developed in the past two decades including calcium phosphate coprecipitation micro injection recombinant viruses liposome-mediated gene transfer lipids that do not form liposomes high molecular weight cationic polymers .