摘要
[目的]以小鼠骨髓瘤(SP2/0)细胞作为实验材料,比较了SP2/0细胞在不同微秒(μs)脉冲和微纳秒(μs/ns)复合脉冲电融合的细胞存活率和融合率,并确定了最优的μs/ns复合脉冲电融合参数。[方法]将SP2/0细胞分别用Hoechst 33342和碘化丙啶(PI)进行染色,分别选取3组μs脉冲和3组μs/ns复合脉冲对染色后的SP2/0细胞进行电融合,通过荧光显微镜观察融合后细胞的存活率和融合率。[结果]对比不同μs和μs/ns复合脉冲电融合存活率和融合率结果得知,μs/ns复合脉冲比μs脉冲电融合效果更好,且电融合参数为2.5 k V/cm、20μs、1次,10 k V/cm、200 ns、8次时存活率和融合效率相对最好,存活率和融合率分别为88.5%±6.2%和80.1%±2.6%。[结论]通过采用不同μs和μs/ns复合脉冲电融合比较了SP2/0细胞的存活率和融合效率,结果显示μs/ns复合脉冲电融合参数在2.5 k V/cm、20μs、1次,10 k V/cm、200 ns、8次时,存活率比μs脉冲电融合提高6.4%,融合率提高23.3%,为利用μs/ns复合脉冲研究杂交瘤细胞电融合奠定了基础。
[Objective]Using mouse myeloma(SP2/0)cells as experimental materials,the survival rate and fusion rate of SP2/0 cells fused with different microsecond(μs)pulses and microsecond/nanosecond(μs/ns)pulses were compared,and the optimal parameters ofμs/ns pulsed electrofusion were determined.[Method]SP2/0 cells were stained with Hoechst 33342 and propidium iodide(PI)respectively.Three groups ofμs pulses and three groups ofμs/ns compound pulses were selected to electrofuse the stained SP2/0 cells.The survival rate and fusion rate of the fused cells were observed by fluorescence microscopy.[Result]By comparing the survival rate and fusion rate of differentμs andμs/ns composite pulse electric fusion,the results show that the composite pulses ofμs/ns are better than those ofμs pulses,and the survival rate and fusion efficiency are the best when the parameters of fusion are 2.5 kV/cm,20μs,1 time,10 kV/cm,200 ns and 8 times,the survival rate and fusion rate are 88.5%±6.2%and 80.1%±2.6%,respectively.[Conclusion]The survival rate and fusion efficiency of SP2/0 cells were compared by using differentμs andμs/ns composite pulsed electrofusion,the results showed that when the parameters ofμs/ns composite pulse electrofusion were 2.5 kV/cm,20μs,once,10 kV/cm,200 ns and 8 times,the survival rate and fusion rate ofμs composite pulse electrofusion were 6.4%and 23.3%higher than those ofμs composite pulse electrofusion,which laid a foundation for the further study of hybridoma cell electrofusion usingμs/ns composite pulse.
作者
吴梦
柯强
刘雪芹
肖普英
李成祥
葛良鹏
WU Meng;KE Qiang;LIU Xue-qin;XIAO Pu-ying;LI Cheng-xiang;GE Liang-peng(Chongqing Academy of Animal Sciences,Chongqing 402460,China;Key Laboratory of Pig Industry Sciences,Ministry of Agriculture,Chongqing 402460,China;Chongqing Key Laboratory of Pig Industry Sciences,Chongqing 402460,China;Chongqing Medical Animal Resources Development and Transformation Application Engineering Technology Center,Chongqing 402460,China;School of Electrical Engineering,Chongqing University,Chongqing 400044,China)
出处
《生物技术》
CAS
2019年第6期551-557,共7页
Biotechnology
基金
重庆市科研院所绩效激励引导专项(cstc2018jxjl0043)
国家自然科学基金面上项目(5167070727)
重庆市农发资金项目(17406)
关键词
微秒
纳秒
复合脉冲
电融合
骨髓瘤细胞
microsecond
nanosecond
compound pulse
electrofusion
myeloma cells
