Satyabhama S, Epstein A L
Department of Pathology, USC Medical Center, Los Angeles 90033.
DNA. 1988 Apr;7(3):203-9. doi: 10.1089/dna.1988.7.203.
The efficiency of DNA transfer into human hematopoietic cells by electroporation was investigated and compared to conventional transfection procedures. Important parameters of electroporation were optimized in human erythroleukemia cells using the chloramphenicol acetyltransferase (acetyl-CoA; chloramphenicol 3-O-acetyltransferase, EC 2.3.1.28) gene linked to the cytomegaloviral enhancer-promoter. In addition, selected chemicals with different modes of action were studied for their ability to aid DNA entry and gene expression in this system, and several were found to enhance gene transfection by electroporation in a significant manner. Using these chemical stimulators, many but not all human and mouse suspension cultures tested were successfully electroporated by the Baekon 2000 instrument. From these studies, it appears that electroporation can be enhanced by chemical additives. Because of its efficiency, reproductivity, and convenience electroporation is an attractive method of gene transfer in human hematopoietic cells.
研究了通过电穿孔将DNA导入人造血细胞的效率，并与传统转染方法进行了比较。使用与巨细胞病毒增强子-启动子相连的氯霉素乙酰转移酶（乙酰辅酶A；氯霉素3-O-乙酰转移酶，EC 2.3.1.28）基因，在人红白血病细胞中优化了电穿孔的重要参数。此外，研究了具有不同作用方式的特定化学物质在该系统中帮助DNA进入和基因表达的能力，发现其中几种能显著增强电穿孔介导的基因转染。使用这些化学刺激剂，通过Baekon 2000仪器成功地对许多（但不是全部）测试的人和小鼠悬浮培养物进行了电穿孔。从这些研究来看，化学添加剂似乎可以增强电穿孔。由于其效率、可重复性和便利性，电穿孔是一种在人造血细胞中进行基因转移的有吸引力的方法。
