Method to produce cloned embryos and adults from cultured cells
OwnerAnthony C.F. Perry, Peter Mombaerts +1inventors
InventorsAnthony C.F. Perry, Peter Mombaerts, Teruhiko Wakayama
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Abstract
A nuclear transfer method is provided wherein nuclear DNA in whole or part is injected into enucleated oocytes. The method is suitable for different donor cells, and preferably ES cells.
Claims72
A method for cloning an embryo comprising the steps of:
- (a) collecting the nucleus of a cultured cell;
- (b) microinjecting the nucleus of (a) or at least a portion of thereof that includes the chromosomes, into an enucleated oocyte to reconstitute the cell;
- and
- (c) allowing the reconstituted cell to develop embryonically.
The method of claim 15, wherein the mammal is a pig.
A method for clonally deriving differentiated cells comprising the steps of:
- (a) collecting the nucleus of an ES cell;
- (b) microinjecting at least a portion of the ES cell nucleus that includes the chromosomes into an enucleated oocyte to form a reconstituted cell;
- (c) incubating the reconstituted cell for 0-6 hours prior to activation;
- (d) activating development of the reconstituted cell;
- and
- (e) allowing the reconstituted cell to develop.
A method for clonally deriving differentiated cells comprising the steps of:
- (a) collecting the nucleus of a cell;
- (b) microinjecting at least a portion of the cell nucleus of (a) that includes the chromosomes into an enucleated oocyte to form a reconstituted cell;
- (c) allowing the reconstituted cell to develop into a morula/blastocyst;
- (d) collecting an ES cell;
- (e) introducing the ES cell of (d) into the morula/blastocyst of (c);
- (f) allowing the reconstituted embryo of (e) to develop.
A method for modulating embryological development, comprising the steps of:
- (a) combining a nucleus of an ES cell with an enucleated oocyte to form a reconstituted cell;
- (b) inserting a reagent into the cytoplasm of the oocyte, prior to, during, or after the combining step;
- and
- (c) allowing the reagent-treated reconstituted cell to develop.
The method of claim 68, wherein the microinjection is piezo electrically-actuated microinjection.
Cells produced by the method of 70.
Dependencies are inferred from the claim text.