鱼类细胞电融合条件的研究

The Study on the Condition of the Electrofusionof Fish Cells

1988—1989年,以草鱼(Ctenopharynoodon idellus)尾鳍组织HGC—87细胞为材料,应用自制BP—88型细胞电融合仪,对鱼类细胞的电融合条件进行研究.草鱼细胞电融合时,需要较高的脉冲场强,低于4KV/cm时,细胞很少发生融合.场强为10KV/cm脉宽60—100μs,连续3—7个脉冲,融合率可达80％,融合后存活率达85—90％。电场介质为3％甘露醇液,含一定浓度的CaCl_2和MgCl_2,Mg^(++)在1mmol/L浓度以下,能促进细胞的极化,但融合时细胞易被电击击破.Ca^(++)在1mmol/L浓度以下,有利于细胞极化成串.融合细胞经21天培养后仍存活,有细胞分裂.

During 1988-1989, the condition of the cell electrofusion of Grass carp (Ctenopharyngodon idellus) HGC-87 cell was studied by means of self -made model BP-88 cell electrofusion apparatus. The fusion of HGC-87 cell must be provided with a higher pulse intensity. When the intensity was less than 4KV/cm, the cell fusion would occur rarely.And when the pulse was 10KV/cm, 60-100μs duration with 3-7 beats, the rate of fusion would reach over 80%, and the rate of vitality of fused-cells would be 85-90%. The electrofusion medium was 3% manintol, containing CaCl2 or MgCl2 at different concentrations. The Mg2+ and Ca2+ could promote the cell's polarization at a suitable concentration, but the Mg2+ could make the cell broken easily by electro-pulses shock. After having cultured 3 weeks, the fused-cells grew normally and appeared mitoses.