摘要
传统的细胞电融合技术通常采用持续时间为微秒级的短时高压电脉冲诱导细胞电穿孔来实现。然而,高幅值微秒脉冲在促进细胞电融合的同时易导致细胞过度穿孔而死亡,从而限制了细胞的融合效率。为进一步提高细胞电融合率,该文基于纳秒脉冲电场所诱导的“超穿孔现象”,结合常规的微秒脉冲电融合技术,提出纳/微秒复合脉冲协同诱导细胞电融合的方法。以SP2/0小鼠骨髓瘤细胞为实验对象,利用COMSOL软件分析复合脉冲作用下细胞膜上跨膜电压与穿孔密度的分布,并进行初步的电融合实验,探究该融合方法对细胞融合率和死亡率的影响。仿真结果显示,传统微秒脉冲作用下,细胞膜的两极与接触区域的跨膜电压均超过穿孔阈值,显著穿孔面积超过10%。而等剂量的纳/微秒复合脉冲作用下,跨膜电压仅在细胞接触区域达到较高值,电穿孔也集中于该区域。实验结果同样证明,该方法能够保证较低细胞死亡率(10%)的同时达到较高的细胞融合效率(75%)。该研究为细胞电融合技术的进一步发展提供了新的思路。
Cell electrofusion technology is usually performed by short-term high-voltage electrical pulses with a duration of microseconds to induce cell electroporation.However,exposure to high amplitude microsecond pulses can easily lead to excessive cell electroporation while promoting cell electrofusion,resulting in the death of the fused cells,thereby limiting the efficiency of cell fusion.To increase the rate of cell electrofusion,this paper proposes a method of synergistic induction of cell electrofusion by combining nanosecond and microsecond pulsed electric fields,based on the phenomenon of“super-electroporation”induced by nanosecond pulsed electric field.In this paper,SP2/0 mouse myeloma cells are used as the experimental object.The COMSOL software is used to analyze the distribution of transmembrane voltage and electroporation density on the cell membrane under the exposure of the composite pulse,and preliminary electrofusion experiments are performed to explore the mortality and fusion rate of the fusion method.The simulation results show that under the traditional microsecond pulse,the transmembrane voltage across the two poles of the membrane and the contact area both exceed the perforation threshold,and the significant perforation area exceeds 10%.Besides,under the action of the same dose of nano/microsecond composite pulse,the transmembrane voltage only reaches a higher value in the cell contact area,and the electroporation is also concentrated in this area.Experimental results also prove that this method can guarantee a lower mortality rate(10%)while achieving a higher cell fusion efficiency(75%),which provides new ideas for the further development of cell electrofusion technology.
作者
李成祥
李新皓
柯强
姚成
姚陈果
Li Chengxiang;Li Xinhao;Ke Qiang;Yao Cheng;Yao Chenguo(State Key Laboratory of Power Transmission Equipment&System Security and New Technology Chongqing University,Chongqing 400030 China)
出处
《电工技术学报》
EI
CSCD
北大核心
2022年第6期1522-1532,共11页
Transactions of China Electrotechnical Society
基金
国家自然科学基金专项基金(81727802)
国家自然科学基金面上(51677017)资助项目。
关键词
细胞电融合
纳/微秒复合脉冲
融合效率
跨膜电压
穿孔密度
Cell electrofusion
nano-microsecond pulsed electric fields
fusion efficiency
transmembrane voltage
electroporation density
