Method for selecting cell lines to be used for nuclear transfer in mammalian species
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Abstract
The present invention provides data to demonstrate that the fusion performance of a cell-line in procedures involving fusion and cleavage indices either alone or in combination are a means for selecting a cell lines that will be successful in a nuclear transfer or microinjection program. This technique and method of selecting a cell line offers an additional alternative and improvement in the creation of activated and fused nuclear transfer-capable embryos for the production of live offspring in various mammalian non-human species including goats, pigs, rodents, primates, rabbits and cattle.
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Citations
48 Claims
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1-23. -23. (canceled)
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24. A method for producing cultured inner cell mass cells, comprising:
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(i) obtaining desired differentiated mammalian cells to be used as a source of donor nuclei;
(ii) obtaining at least one oocyte from a mammal of the same species as the cells which are the source of donor nuclei;
(iii) enucleating said at least one oocyte;
(iv) transferring the desired differentiated cell or cell nucleus into the enucleated oocyte;
(v) simultaneously fusing and activating the cell couplet to form a first transgenic embryo;
(vi) activating a cell-couplet to create a first transgenic embryo that is activated after an initial electrical shock; and
(vi) culturing cells obtained from said cultured activated embryo to obtain cultured inner cell mass cells;
(vii) wherein the desired differentiated mammalian cell line to be used as a karyoplast is selected according to the objective parameters of cleavage and/or fusion patterns - View Dependent Claims (25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47)
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48-53. -53. (canceled)
Specification