Biotechnology Science and Engineering, University of Alabama in Huntsville, Huntsville, AL, USA.
Department of Chemical and Materials Engineering, University of Alabama in Huntsville, Huntsville, AL, USA.
Drug Deliv. 2020 Dec;27(1):805-815. doi: 10.1080/10717544.2020.1770371.
Jurkat, an immortalized cell line derived from human leukemic T lymphocytes, has been employed as an excellent surrogate model of human primary T-cells for the advancement of T-cell biology and their applications in medicine. However, presumably due to its T-cell origin, Jurkat cells are very difficult to transfect. Thus, for the genetic modification of Jurkat cells, expensive and time-consuming viral vectors are normally required. Despite many previous efforts, non-viral vectors have not yet overcome the hurdles of low transfection efficiency and/or high toxicity in transfection of Jurkat cells. Here, we report that a simple addition of calcium ions (Ca) into culture media at optimal concentrations can enhance the efficiency of the polyplex-mediated transfection using poly(ethylene imine) (PEI) by up to 12-fold when compared to the polyplex-only control. We show that calcium enhances the association between polyplex and Jurkat, which is at least partially responsible for the increase in transmembrane delivery of polyplex and consequential enhancement in expression of transgene. Other cations, Mg or Na did not show similar enhancement. Interestingly, addition of Ca was rather detrimental for the transfection of lipoplex on Jurkat cells. Observation of significant enhancement in the transfection of non-viral vectors with a simple and physiologically relevant reagent like Ca in the engineering of hard-to-transfect cells such as Jurkat warrants further investigation on similar strategies.
Jurkat 是一种源自人类白血病 T 淋巴细胞的永生化细胞系，已被用作人类原代 T 细胞的优秀替代模型，用于推进 T 细胞生物学及其在医学中的应用。然而，由于 Jurkat 细胞来源于 T 细胞，因此它非常难以转染。因此，对于 Jurkat 细胞的基因修饰，通常需要昂贵且耗时的病毒载体。尽管之前进行了许多努力，但非病毒载体尚未克服在转染 Jurkat 细胞时转染效率低和/或毒性高的障碍。在这里，我们报告称，在最佳浓度下向培养基中添加钙离子（Ca）可以将聚电解质复合物（PEI）介导的转染效率提高多达 12 倍，与仅使用聚电解质复合物的对照相比。我们表明，钙离子增强了聚电解质复合物与 Jurkat 之间的结合，这至少部分解释了聚电解质复合物跨膜传递的增加以及随后转基因表达的增强。其他阳离子，如镁或钠，没有显示出类似的增强作用。有趣的是，添加 Ca 对 Jurkat 细胞中脂质体的转染反而有害。在难以转染的细胞（如 Jurkat）的工程中，使用 Ca 等简单且与生理相关的试剂对非病毒载体的转染进行显著增强，这值得进一步研究类似的策略。
