Patent ID: 9546383
Date: 2017-01-17
CPC Classifications: C12N

Claim:
1. A method of producing a human pluripotent embryonic stem cell, the method comprising: enucleating a human oocyte from a donation cycle of 15 or fewer oocytes by removing the MII spindle in a manner that does not lower levels of maturation promoting factor, thereby producing a cytoplast; contacting a human donor nucleus with an HVJ-E extract; contacting the cytoplast with the human donor nucleus thereby producing an SCNT embryo; treating the human oocyte and/or the cytoplast and/or the donor nucleus and/or the SCNT embryo with a protein phosphatase inhibitor; applying a first electroporation pulse to the SCNT embryo, thereby producing an activated SCNT embryo; culturing the activated SCNT embryo in a first media comprising 6-DMAP; culturing the activated SCNT embryo in a second media comprising TSA; culturing the activated SCNT embryo in a third media thereby producing a blastocyst; culturing the blastocyst on a feeder layer; and selecting a cell with an embryonic stem cell-like morphology.