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US 5,843,754 AGranted
Parthenogenic bovine oocyte activation
AssigneeABS Global, Inc.
InventorsJoan L. Susko-Parrish, David L. Northey, M. Lorraine Leibfried-Rutledge, Steven L. Stice
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Abstract
A process of parthenogenic activation of mammalian oocytes which includes increasing intercellular levels of divalent cations in the oocyte; and reducing phosphorylation of cellular proteins in the oocyte. One method of accomplishing this is by introducing Ca.sup.2+ free cation, such as ionomycin, to the oocyte and then preventing phosphorylation of the cellular proteins within the oocyte by adding a serine-threonine kinase inhibitor, such as 6-dimethylaminopurine (DMAP).
Claims19
A method for multiplying bovine embryos in vitro comprising:
- a. collecting bovine embryonic cells;
- b. isolating individual bovine embryonic cells;
- c. culturing the individual bovine embryonic cells;
- d. collecting recipient bovine oocytes from donor bovine animals in vitro;
- e. enucleating the recipient bovine oocytes:
- f. parthenogenically activating the recipient bovine oocytes, wherein the bovine oocytes are activated by a process comprising the following steps in sequence:
- i. increasing intracellular levels of divalent cations in the bovine oocyte;
- and
- ii. reducing phosphorylation of cellular proteins in the bovine oocyte;
- g. transferring the nucleus from the cultured bovine embryonic cell to an enucleated recipient parthenogenically-activated bovine oocyte to form a nuclear transferred oocyte;
- and
- h. forming a single cell bovine embryo from the nuclear transferred bovine oocyte.
9 more elements
Dependencies are inferred from the claim text.