利用改良电融合法制备杂交瘤细胞的试验探索  

The Study of Modified Electrofusion Method to Prepare Hybridoma Cell

作　　者：吴梦[1,2] 刘作华[1,2,3] 罗林[1] 丁玉春[1,2] 葛良鹏[1,2,3] 

机构地区：[1]重庆市畜牧科学院,重庆荣昌402460 [2]农业部养猪科学重点实验室,重庆荣昌402460 [3]养猪科学重庆市市级重点实验室,重庆荣昌402460

出　　处：《四川畜牧兽医》2016年第7期26-28,共3页Sichuan Animal & Veterinary Sciences

基　　金：重庆市基本科研业务费项目(2015cstc-jbky-00120;2014cstc-jbky-00111);国家国际科技合作专项(2013DFA31820);863项目(2014AA021602);重庆市国际合作项目(CSTC2013gjhz80002);重庆市基础与前沿研究重大项目(cstc2013jcyj C80001);重庆市农发资金项目(12402)

摘　　要：目的:运用改良电融合法提高杂交瘤的融合效率,以获得更多的杂交瘤克隆。方法:将SP2/0细胞和脾细胞分别用NHS-d-PEG24-biotin孵育30 min,再将脾细胞用avidin孵育30 min,然后将SP2/0细胞和脾细胞按1∶5混合孵育15 min后进行电融合,融合后的细胞用半固体培养基筛选单克隆。结果显示:用2×105个SP2/0细胞和1×106个脾细胞进行正常电融合,经半固体培养基筛选共获得6个单克隆,而用改良电融合法获得了20个单克隆。结论:利用改良电融合法能够增加异种细胞相互配对的几率,从而提高杂交瘤的融合效率,最终获得更多的杂交瘤细胞。By using the method of modified electrofusion to improve the hybridoma fusion efficiency and obtain more hybridoma clones. SP2/0 cells and spleen cells were incubated with NHS-d-PEG24-biotin for 30min, then spleen ceils were incubated with avidin for 30min respectively, and the SP2/O ceils and spleen cells were incubated by ration of 1：5 in 15 min, the fused cells used semi-solid medium for screening monoclonal. Using 2x105 SP2/0 cells and 1~106 spleen cells for normal electrofusion~ the semi-solid medium screening obtained a total of 6 monoclonals, while modified electrofusion obtained 20 clones. The result showed that modified electrofusion could obviously increase the matching probability of heterogeneous cells, so as to improve the efficiency of hybridoma fusion and obtain more hybridoma cells.

关 键 词：电融合 半固体培养基 杂交瘤 

分 类 号：Q813[生物学—生物工程]