| 摘要 |
<p>PROBLEM TO BE SOLVED: To fuse cells for the production, or the like, of a clone animal or a transgenic animal by treating the cells with a polyethylene glycol solution and a dimethylsulfoxide solution, and fusing the treated cells with an enucleated ovum by an electron pulse method. SOLUTION: Colostrum collected within seven days, preferably one day after parturition is doubly diluted with a sterilized phosphate buffer solution(PBS) at pH 7.0, and the diluted product is subjected to a centrifugal separation to provide a sedimentation. The obtained sedimentation is suspended in a Dolbecco' s modified Eagle's medium containing fetal bovine serum in 15% concentration and an antibiotic in a concentration of 10 times as much as that at the normal time, and cultured at 37 deg.C in the presence of 5% CO2 to provide breast epithelial cells. The cells such as the obtained breast epithelial cells are treated with 10-80% polyethylene glycol(PEG) and 1-20% dimethylsulfoxide(DMSO) solutions, and the treated cells and an enucleated ovum prepared from ovum-ovarian cumulus cell complex collected from a bovine carcass are fused by an electron pulse method to provide the objective fused cell useful for production, or the like, of a clone animal, a transgenic animal, or the like.</p> |
