Chang D C, Gao P Q, Maxwell B L
Department of Molecular Physiology and Biophysics, Baylor College of Medicine, Houston, TX 77030.
Biochim Biophys Acta. 1991 Apr 17;1092(2):153-60. doi: 10.1016/0167-4889(91)90149-r.
In order to develop a safe and effective way to introduce exogenous genes into cells, we have experimented with a new method of electroporation which uses a radio-frequency (RF) electric field to permeabilize the cell membrane. This RF method has several advantages over the conventional electroporation method which uses a direct current (DC) field. We have shown that the RF electroporation method can be used to introduce marker genes into a wide variety of cell lines, including COS-M6, CV-1, CHO, 3T3 and hepatocytes, and is able to increase substantially the efficiency of gene transfection. (For example, the amount of DNA required for transfecting two million COS-M6 cells can be as low as 0.1 microgram). The transfection efficiency is shown to be affected by a number of factors, including cell type, field strength, pulse protocol and medium buffer. Because of its wide range of applications, high transfection efficiency and lack of harmful side-effect, the RF electroporation method would be particularly useful for introducing genes into human cells for gene therapy.
为了开发一种将外源基因导入细胞的安全有效的方法，我们试验了一种新的电穿孔方法，该方法使用射频（RF）电场使细胞膜通透性增加。与使用直流（DC）电场的传统电穿孔方法相比，这种射频方法具有几个优点。我们已经表明，射频电穿孔方法可用于将标记基因导入多种细胞系，包括COS-M6、CV-1、CHO、3T3和肝细胞，并且能够大幅提高基因转染效率。（例如，转染两百万个COS-M6细胞所需的DNA量可低至0.1微克）。转染效率受多种因素影响，包括细胞类型、场强、脉冲方案和培养基缓冲液。由于其广泛的应用范围、高转染效率以及无有害副作用，射频电穿孔方法对于将基因导入人体细胞进行基因治疗将特别有用。
