Embryonic or stem-like cell lines produced by cross species...

C - Chemistry – Metallurgy – 12 – N

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C12N 15/87 (2006.01) A61K 35/12 (2006.01) A61K 35/14 (2006.01) C12N 5/10 (2006.01) C12N 5/08 (2006.01)

Patent

CA 2384413

An improved method of nuclear transfer involving the transplantation of differentiated 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 therapied and for in vitro study of cell differentiation. Also, methods for improving nuclear transfer efficiency by genetically altering donor cells to inhibit apoptosis, select for a specific cell cycle and/or enhance embryonic growth and development are provided.

La présente invention concerne une technique améliorée de transfert nucléaire consistant à transplanter des noyaux de cellules donneuses différenciées dans des oocytes énucléés d'espèces différentes de celle de la cellule donneuse. Les unités de transfert nucléaire qui en résultent sont utiles pour la production de cellules souches embryonnaires isogéniques, en particulier des cellules souches embryonnaires isogéniques humaines. Ces cellules embryonnaires ou de type souche conviennent pour la production de cellules différenciée recherchées et pour introduire, retirer ou modifier des gènes recherchés, par exemple au niveau de sites spécifiques du génome de ces cellules par recombinaison homologue. Ces cellules, qui peuvent contenir un gène hétérologue, conviennent particulièrement pour les thérapies par transplantation cellulaire et pour les études in vitro de différenciation cellulaire. Cette invention concerne aussi des techniques permettant d'améliorer l'efficacité du transfert nucléaire par modification génétique de cellules donneuses de façon à inhiber l'apoptose, à sélectionner un cycle cellulaire spécifique et/ou améliorer la croissance et le développement embryonnaire.

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