Department of Anesthesiology, Chang Gung Memorial Hospital-Kaohsiung Medical Center, Chang Gung University College of Medicine, Kaohsiung, Taiwan.
J Biomed Sci. 2010 Jun 3;17(1):44. doi: 10.1186/1423-0127-17-44.
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.
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.
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%.
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.
基因转移已成功应用于许多细胞类型，以开发替代和辅助传统医学治疗的方法。然而，有丝分裂后神经元的有效转染仍然是一个挑战。本研究旨在开发一种使用声孔法将基因转移到大鼠原代背根神经节神经元中的方法。
对成年大鼠背根神经节（DRG）细胞的分离细胞进行 1-8 秒的超声处理，以确定用于基因转染和细胞存活的最佳超声持续时间和功率。比较了声孔法、脂质体和慢病毒载体基因转移技术之间的转染效率。
最佳超声强度为 5 W 持续 2 秒，基因转染效率为 31%，存活率为 35%。
可以优化声孔法以最小化细胞死亡并获得高比例的转染神经元，并且该技术可以很容易地应用于大鼠背根神经节神经元的原代培养。
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