Sonoporation-mediated gene transfer into adult rat dorsal root ganglion cells.
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 2.5-10 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.
Department of Anesthesiology, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung, Taiwan. chungren@ntu.edu.tw
Chung-Ren
CR
Lin
Kuan-Hung
KH
Chen
Chien-Hui
CH
Yang
Jiin-Tsuey
JT
Cheng
Shyr-Ming
SM
Sheen-Chen
Chih-Hsien
CH
Wu
Wei-Dih
WD
Sy
Yi-Shen
YS
Chen
D000818:Animals; D002470:Cell Survival; D002478:Cells, Cultured; D005727:Ganglia, Spinal / Q000166:cytology* / Q000378:metabolism*; D018014:Gene Transfer Techniques; D005822:Genetic Vectors; D049452:Green Fluorescent Proteins / Q000235:genetics; D016086:Lentivirus / Q000235:genetics; D008081:Liposomes; D009474:Neurons / Q000166:cytology / Q000378:metabolism; D051381:Rats; D011994:Recombinant Proteins / Q000235:genetics; D014162:Transfection; D014404:Tubulin / Q000378:metabolism; D014465:Ultrasonics
D008081:Liposomes; D011994:Recombinant Proteins; D014404:Tubulin; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins