Ohyama H, McBride J, Wong D T
Department of Oral Medicine and Diagnostic Sciences, Harvard School of Dental Medicine, Boston, MA 02115, USA.
J Immunol Methods. 1998 Jun 1;215(1-2):105-11. doi: 10.1016/s0022-1759(98)00067-2.
Eosinophils are emerging as an increasingly important cell in the immunoregulatory network of normal and pathological processes. No studies has yet described optimized experimental strategies to transfect DNA into human eosinophils. Using a frequently employed in vitro model of human eosinophil, the EoL-1 cells, we now described the optimal transfection of DNA into these cells by electroporation. Our results indicate that electroporation can efficiently and reproducibly transfect DNA into EoL-1 cells. Optimal electroporation conditions consist of the use of 1 X RPMI medium 1640 with 10% FBS, voltage setting at 275 V, 1150 microF capacitance, 40 mg of DNA and 4.0 X 10(7) cells/ml per electroporation in a total volume of 0.5 ml in 0.4 cm gap cuvettes. These conditions may be a useful protocol for transfecting eosinophil cell lines.
嗜酸性粒细胞在正常和病理过程的免疫调节网络中逐渐成为一种越来越重要的细胞。尚无研究描述将DNA转染入人嗜酸性粒细胞的优化实验策略。我们利用一种常用的人嗜酸性粒细胞体外模型EoL-1细胞，描述了通过电穿孔将DNA最佳转染入这些细胞的方法。我们的结果表明，电穿孔可有效且可重复地将DNA转染入EoL-1细胞。最佳电穿孔条件包括使用含10%胎牛血清的1×RPMI 1640培养基，电压设置为275 V，电容为1150 μF，每次电穿孔使用40 μg DNA和4.0×10⁷个细胞/ml，总体积为0.5 ml，使用0.4 cm间隙的比色杯。这些条件可能是转染嗜酸性粒细胞系的有用方案。
