Method for producing enucleated and nucleated fragments of...

A - Human Necessities – 61 – D

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A61D 19/04 (2006.01)

Patent

CA 2055247

ABSTRACT OF THE DISCLOSURE A method for producing enucleated and nucleated membrane-bounded egg fragments from ovulated or in vitro matured mammalian eggs, includes the following steps: 1) removal of any cells attached to the zona pellucida. 2) opening or removal of the zona pellucida. 3) incubation of the denuded egg for about an hour in a saline medium of 290 mosm and near neutral pH for visualization of the location of the chromosomes, and 4) mechanical division of the egg into one membrane bounded fragment containing the chromosomes, and one or more membrane bounded anucleate fragments. The size of nucleated and enucleated fragments may be varied at will within the limits set by the size of the egg at the outset and a) - in the case of the nucleated fragment - the minimum size necessary to accomodate the chromosomes within it, or b) - in the case of enucleated fragment(s) the minimum size obtainable with the instruments used to produce the particular fragment. The enucleated and nucleated egg fragments may be used for the production of composite eggs and embryos by nuclear and cytoplasmic transplantation.

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