Rems Lea, Ušaj Marko, Kandušer Maša, Reberšek Matej, Miklavčič Damijan, Pucihar Gorazd
1] University of Ljubljana, Faculty of Electrical Engineering, Tržaška cesta 25, SI-1000 Ljubljana, Slovenia [2].
Sci Rep. 2013 Nov 29;3:3382. doi: 10.1038/srep03382.
Electrofusion is an efficient method for fusing cells using short-duration high-voltage electric pulses. However, electrofusion yields are very low when fusion partner cells differ considerably in their size, since the extent of electroporation (consequently membrane fusogenic state) with conventionally used microsecond pulses depends proportionally on the cell radius. We here propose a new and innovative approach to fuse cells with shorter, nanosecond (ns) pulses. Using numerical calculations we demonstrate that ns pulses can induce selective electroporation of the contact areas between cells (i.e. the target areas), regardless of the cell size. We then confirm experimentally on B16-F1 and CHO cell lines that electrofusion of cells with either equal or different size by using ns pulses is indeed feasible. Based on our results we expect that ns pulses can improve fusion yields in electrofusion of cells with different size, such as myeloma cells and B lymphocytes in hybridoma technology.
电融合是一种利用短时长高压电脉冲来融合细胞的高效方法。然而，当融合伙伴细胞的大小差异很大时，电融合的产量非常低，因为使用传统微秒脉冲时的电穿孔程度（进而膜融合状态）与细胞半径成正比。我们在此提出一种全新的创新方法，即使用更短的纳秒（ns）脉冲来融合细胞。通过数值计算，我们证明纳秒脉冲能够诱导细胞间接触区域（即目标区域）的选择性电穿孔，而与细胞大小无关。然后，我们在B16 - F1和CHO细胞系上通过实验证实，使用纳秒脉冲对大小相同或不同的细胞进行电融合确实可行。基于我们的结果，我们预计纳秒脉冲能够提高不同大小细胞电融合的产量，比如在杂交瘤技术中骨髓瘤细胞和B淋巴细胞的融合。
