›› 2009, Vol. 29 ›› Issue (6): 366-371.

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Effect of Fusion Conditions on Efficiency of Electrofusion of Mouse Somatic Cell Nuclear Transfer

  

  1. 1. The Comparative Medicine Center of Yangzhou University, Yangzhou 225009, China 2. College of Veterinary Medicine, Yangzhou University, Yangzhou 225009, China
  • Received:2009-05-25 Online:2009-06-30 Published:2009-06-30

Abstract: Objective To improve the efficiency of electrofusion of reconstructed embryos derived from oocytes and fetal fibroblast cells both of B6D2F1strain within a mouse somatic cell nuclear transfer(SCNT)program.Methods The effect of different pre-treatment on donor cells,fusion midia,electrical field intensity,electrodes’shape and cell passage on the efficiency of electro fusion was studied.Results(l)The pre-treatment of PEG/DMSO on donor cells significantly improved the fusion rate(51.11%vs 25.64%)(P<0.05);(2)Fusion rate in the fusion media containing O.lg/L polyvinyl alcohol(PVA)was higher than those in the media without PVA.Fusion rate in fusion media containing 0.32 mol/L mannitol(48.21%)was higher than those in the media with 0.28 mol/L,0.30 mol/L and 0.36 mol/L mannitol(27.78%,41.67%and 30.11%);(3)Although the chamber is a much more routine way for fusing the reconstructed embryos,the tip shape electrodes can improve the fusion rate(42.31%,43.13%)comparing to routine parallel line electrodes(30.43%,33.33%);(4)the efficiency of electrofusion of oocytes with fetal fibroblast cells at the found passage and the first passage was higher than that at the second and the third passage(38.98%,38.46%,36.73%and 35.00%),regardless of other fusion conditions.Conclusion Both the pre-treatment of PEG/DMSO on donor cells and the addition of PVA to the fusion media can improve the efficiency of electro fusion in the procedure of mouse SCNT.

Key words: PEG, Fusion, Mouse, Somatic cell nuclear transfer