Schoenberg Kathrin, Trompeter Hans-Ingo, Uhrberg Markus
University Clinic of Düsseldorf, Institute for Transplantation Diagnostics and Cell Therapeutics, Düsseldorf, Germany.
Methods Mol Biol. 2008;423:165-72. doi: 10.1007/978-1-59745-194-9_11.
Natural killer (NK) cells are highly resistant to transfection by conventional methods such as electroporation and lipofection. Recently, we reported the employment of a novel electroporation-based method, called nucleofection, which for the first time enabled efficient nonviral gene transfer into NK cells. In this study, we aimed at developing optimized conditions for the transfection of different NK cell lines as well as primary NK cells. Using EGFP (enhanced green fluorescent protein) or luciferase as reporter genes, suitable buffer conditions as well as instrument settings were defined. The new transfection methodology represents a useful tool for the immunotherapeutic use of NK cells, with the potential to enhance cytotoxicity as well as retarget the specificity of cytotoxic lymphocytes in clinical therapy of cancer and viral infection.
自然杀伤（NK）细胞对诸如电穿孔和脂质转染等传统方法的转染具有高度抗性。最近，我们报道了一种基于电穿孔的新方法，称为核转染，它首次实现了将非病毒基因高效导入NK细胞。在本研究中，我们旨在为不同NK细胞系以及原代NK细胞的转染开发优化条件。使用增强型绿色荧光蛋白（EGFP）或荧光素酶作为报告基因，确定了合适的缓冲液条件以及仪器设置。这种新的转染方法是NK细胞免疫治疗应用的一种有用工具，有可能增强细胞毒性，并在癌症和病毒感染的临床治疗中重新靶向细胞毒性淋巴细胞的特异性。
