Embryonic or stem-like cell lines produced by cross species nuclear transplantation
First Claim
1. A method of producing embryonic or stem-like cells comprising the following steps:
- (i) inserting a desired human or mammalian cell or cell nucleus into an enucleated animal oocyte, wherein such oocyte is derived from a different animal species than the human or mammalian cell under conditions suitable for the formation of a nuclear transfer (NT) unit;
(ii) activating the resultant nuclear transfer units;
(iii) culturing said activated nuclear transfer units until greater than the 2-cell developmental stage; and
(iv) culturing cells obtained from said cultured NT units to obtain embryonic or stem-like cells.
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Abstract
An improved method of nuclear transfer involving the transplantation of donor cell nuclei into enucleated oocytes of a species different from the donor cell is provided. The resultant nuclear transfer units are useful for the production of isogenic embryonic stem cells, in particular human isogenic embryonic or stem cells. These embryonic or stem-like cells are useful for producing desired differentiated cells and for introduction, removal or modification, of desired genes, e.g., at specific sites of the genome of such cells by homologous recombination. These cells, which may contain a heterologous gene, are especially useful in cell transplantation therapies and for in vitro study of cell differentiation.
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Citations
35 Claims
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1. A method of producing embryonic or stem-like cells comprising the following steps:
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(i) inserting a desired human or mammalian cell or cell nucleus into an enucleated animal oocyte, wherein such oocyte is derived from a different animal species than the human or mammalian cell under conditions suitable for the formation of a nuclear transfer (NT) unit;
(ii) activating the resultant nuclear transfer units;
(iii) culturing said activated nuclear transfer units until greater than the 2-cell developmental stage; and
(iv) culturing cells obtained from said cultured NT units to obtain embryonic or stem-like cells. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35)
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Specification