METHOD FOR ACTIVATING FUSION CELL TO PRODUCE SOMATIC CELL CLONED ANIMALS

摘要

PURPOSE: A method for activating a fusion cell is provided for producing somatic cell hybrids, that is cloned animals, so that animals can be effectively cloned by the somatic cell hybridization technique. CONSTITUTION: The method for activating a fusion cell comprises the steps of: collecting immatured oocyte and incubating; isolating somatic cells from the various tissues of a cow and storing; cutting partially the oolemma of the matured oocyte, squeezing the oocyte and removing 10 to 30% of cytoplasm to remove nucleus from oocyte; transplanting the isolated somatic cells into the enucleated oocyte; electrofusing the squeezed and transplanted oocyte at a voltage under DC condition; and activating the fusion cell in a medium containing chemicals; culturing the activated cell in a postactivation medium and a micro-droplet medium; and assaying the oocyte. The transplanted oocyte is simultaneously electrofused and activated in a medium containing Ca2+, and the transplanted oocyte is also activated by an ionomycin treatment method which is carried out by storing the fusion cell in 50 microliter well for 4 minutes without lighting, removing ionomycin in washing solution added with 10% FBS or 30mg/ml BSA in 35 mm dish, and storing 5 minutes.