Generation of human oocytes
AssigneeOregon Health & Science University
InventorsShoukhrat Mitalipov, Nuria Marti-Gutierrez, Dmitry Temiakov
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Abstract
Methods are provided of generating functional human oocytes following nuclear transfer of first polar body (PB1) genomes from metaphase II (MII) oocytes into enucleated donor MII cytoplasm (PBNT) and using mitochondrial replacement techniques to circumvent mother-to-child mtDNA disease transmission.
Claims17
A method for preparing a viable human oocyte for fertilization, the method comprising the steps of:
- a) isolating a first oocyte containing a polar body;
- b) gaining access through the zona pellucida of the first oocyte;
- c) removing the polar body from the first oocyte;
- d) removing the nuclear spindle from a second oocyte to create an enucleated cytoplast;
- e) placing the polar body removed from the first oocyte into the perivitelline space of the enucleated cytoplast, thereby producing the viable human oocyte.
A method of producing a human oocyte in vitro, the method comprising:
- a) determining a mitochondrial DNA sequence of a conserved sequence box II of a donor oocyte, thereby identifying the conserved sequence box genotype of the donor oocyte;
- b) enucleating the donor oocyte while the donor oocyte is arrested at metaphase II, thereby isolating donor nuclear genetic material;
- c) introducing the donor nuclear genetic material into an enucleated recipient oocyte, wherein mitochondrial DNA of the donor oocyte and mitochondrial DNA of the recipient oocyte have the same conserved sequence box II genotype.
A method of producing a human oocyte in vitro, the produced human oocyte comprising minimal maternal mitochondrial DNA, the method comprising:
- determining the sequence of the major non-coding region of the mitochondrial DNA of a donor oocyte, where the major non-coding region sequence includes a D loop region;
- enucleating the donor oocyte while the donor oocyte is arrested at metaphase II, thereby isolating donor nuclear genetic material;
- introducing the donor nuclear genetic material into an enucleated recipient oocyte, provided that the mitochondrial DNA of the donor oocyte has the same major non-coding region sequence as the recipient oocyte.
Dependencies are inferred from the claim text.