电脉冲介导基因导入悬浮培养哺乳动物细胞的最佳条件

Optimum conditions for electric pulse-mediated gene transfer to mammalian cells in suspension.

作者信息

Hama-Inaba H, Takahashi M, Kasai M, Shiomi T, Ito A, Hanaoka F, Sato K

出版信息

Cell Struct Funct. 1987 Apr;12(2):173-80. doi: 10.1247/csf.12.173.

DOI:10.1247/csf.12.173

Abstract

A pulse-generating machine which delivers exponentially decaying pulses over broad range of pulse lengths was used to determine the optimum pulse conditions for gene transfer to FM3A cells. In the transformation of tk- cells with pTK1, a single pulse of 100-2000 microseconds gave a high transformation frequency at 1.5-6 kV/cm and room temperature, the highest transformation frequency obtained being 3 X 10(-3). As the suspension buffer for cells exposed to the pulse, Saline G was better than PBS(-) for obtaining a large number of transformants because it ensured high cell viability.

摘要

使用一台能在很宽的脉冲长度范围内产生指数衰减脉冲的脉冲发生器，来确定基因转移至FM3A细胞的最佳脉冲条件。在用pTK1转化tk-细胞时，在1.5 - 6 kV/cm和室温下，100 - 2000微秒的单个脉冲可产生较高的转化频率，获得的最高转化频率为3×10(-3)。作为暴露于脉冲的细胞的悬浮缓冲液，生理盐水G比PBS(-)更有利于获得大量转化体，因为它能确保高细胞活力。