METHOD OF ENUCLEATION AND OOCYTE ACTIVATION IN SOMATIC CELL NUCLEAR TRANSFER IN PRIMATES

摘要

METHOD OF ENUCLEATION AND OOCYTE ACTIVATION IN SOMATIC CELLNUCLEAR TRANSFER IN PRIMATES ABSTRACT The present invention relates to a method for cell enucleation, comprisingremoval of the spindle body from a cell in metaphase with a minimal amount of cytoplasm from the cell, said method comprising using polarized light microscopy for visualization of the spindle body during cell enucleation. The present invention also relates to a method for production of an activated reconstructed vertebrate cell, said method comprising: a) culturing a reconstructed vertebrate cell for about 1.5 to about 3 hours after being reconstructed; b) applying at least one electrical pulse to the reconstructed cell; c) culturing the reconstructed cell for about a further 2 hours; and d) treating the reconstructed cell with a chemical activator. The methods of the invention find application in the preparation of totipotent or pluripotent cells of substantially identical genotype, as well as the cloning of vertebrates.(no figure to-accompany the abstract)