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

C - Chemistry – Metallurgy – 12 – N

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C12N 15/87 (2006.01) C12N 5/073 (2010.01) C12N 5/0735 (2010.01) C12N 15/873 (2010.01)

Patent

CA 2471827

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 therapies 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 qui implique la transplantation de noyaux de cellules donneuses différentiées d'ovocytes énucléés d'une espèce différente de la cellule donneuse. Les unités de transfert nucléaire résultantes conviennent pour la production de cellules souche embryonnaires isogéniques, en particulier pour des cellules souches ou des cellules embryonnaires isogéniques humaines. Ces cellules de type souche ou embryonnaires conviennent pour produire des cellules différentiée 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 des thérapies par transplantation de cellules et pour l'étude in vitro de la différentiation cellulaire. Cette invention concerne aussi des techniques permettant d'améliorer l'efficacité du transfert nucléaire par une modification génétique de cellules donneuses de façon à inhiber l'apoptose, sélectionner un cycle cellulaire spécifique et/ou renforcer la croissance et le développement embryonnaire cellulaire.

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