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

C - Chemistry – Metallurgy – 12 – N

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C12N 5/00 (2006.01) C12N 15/00 (2006.01) C12N 15/87 (2006.01) A61K 35/12 (2006.01)

Patent

CA 2323094

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.

L'invention concerne une méthode améliorée de transfert nucléaire, consistant à transplanter des noyaux de cellules donneuses différentiées dans des ovocytes énucléés d'une espèce différente de celle de la cellule donneuse. Les unités de transfert nucléaire ainsi obtenues sont utiles pour produire des cellules souches embryonnaires isogéniques, en particulier des cellules souches ou embryonnaires isogéniques humaines. Ces cellules embryonnaires ou de type souche sont par ailleurs utiles pour produire les cellules différentiées voulues, et pour introduire, éliminer, ou modifier les gènes souhaités, par exemple sur les sites spécifiques du génome de ces cellules, par recombinaison homologue. Ces cellules, qui peuvent renfermer un gène hétérologue, sont particulièrement utiles dans des thérapies de transplantation cellulaire et pour l'étude in vitro de la différentiation cellulaire. L'invention concerne également des méthodes permettant d'améliorer l'efficacité du transfert nucléaire grâce à une modification génétique des cellules donneuses, destinée à inhiber l'apoptose, à choisir un cycle cellulaire spécifique, et/ou à favoriser la croissance et le développemebt embryonnaire.

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