CELL FUSION METHOD FOR PRODUCTION OF SOMATIC CELL CLONED ANIMAL

摘要

PURPOSE: A cell fusion method for production of somatic cell cloned animal is provided to fuse cell after transplantation of donor cell in production of animal like bovine by in vitro fertilization and nuclear transplantation technique. CONSTITUTION: In cell fusion method which is used as production method of cloning bovis, a cell fusion method is characterized by comprising the following steps: (a)preparing a fertile egg or somatic cell of bovine obtained by in vitro fertilization or in vitro culture; (b)separating cell line from various tissue of bovine; (c)performing enucleation by incising part of transparent zone of mature egg prepared in step(a) and removing 10-30% of cytoplasm; (d)transplanting donor cell prepared in step(b) into the egg enucleated in step(c); (e)performing electric fusion of cell transplanted and adhered in step(d) by using 0.75-2.25 kv/cm of DC condition, 1-2 times so as to control the number of times x time within total 60 microsecond; (f)activating the fusion cell in medium; (g)culturing the activated cell in micro-medium after activation and inspection of egg.