Process of mammalian cell reprogramming through production of a heterokaryon

A method of preparing a reprogrammed genetically modified diploid mammalian cell comprising

• providing a genetically modified diploid mammalian donor nucleus derived from a genetically modified somatic cell, and a mammalian recipient cell, wherein the genetically modified diploid mammalian donor nucleus and the mammalian recipient cell are of the same species;
• introducing the genetically modified mammalian donor nucleus into the mammalian recipient cell to produce an aneuploid cell;
• maintaining the aneuploid cell in a suitable environment for a period sufficient to allow the donor nucleus to be reprogrammed;
• subjecting the aneuploid cell to an activation step;
• and
• subsequent to maintaining the aneuploid cell in the suitable environment, treating said reprogrammed aneuploid cell so as to generate a reprogrammed genetically modified diploid mammalian cell from said reprogrammed aneuploid cell by removal or destruction of the mammalian recipient cell nucleus, pronucleus, metaphase plate, chromatin, chromosomes or nuclear DNA from said reprogrammed aneuploid cell.

A method of preparing a reprogrammed genetically abnormal diploid mammalian cell comprising

• providing a genetically abnormal diploid mammalian donor nucleus derived from a genetically abnormal somatic cell comprising a mutation associated with a genetic disease, and a mammalian recipient cell, wherein the genetically abnormal diploid mammalian donor nucleus and the mammalian recipient cell are of the same species;
• introducing the genetically abnormal mammalian donor nucleus into the mammalian recipient cell to produce an aneuploid cell;
• maintaining the aneuploid cell in a suitable environment for a period sufficient to allow the donor nucleus to be reprogrammed;
• subjecting the aneuploid cell to an activation step;
• and
• subsequent to maintaining the aneuploid cell in the suitable environment, treating said reprogrammed aneuploid cell so as to generate a reprogrammed genetically abnormal diploid mammalian cell from said reprogrammed aneuploid cell by removal or destruction of the mammalian recipient cell nucleus, pronucleus, metaphase plate, chromatin, chromosomes or nuclear DNA from said reprogrammed aneuploid cell.

A method of generating one of a group consisting of a cell and cell line from a reprogrammed diploid mammalian cell, comprising:

• (a) preparing a reprogrammed diploid mammalian cell by providing:
• providing a diploid mammalian donor nucleus derived from a somatic cell, and a mammalian recipient cell, wherein the diploid mammalian donor nucleus and the mammalian recipient cell are of the same species;
• introducing the mammalian donor nucleus into the mammalian recipient cell to produce an aneuploid cell;
• maintaining the aneuploid cell in a suitable environment for a period sufficient to allow the donor nucleus to be reprogrammed;
• subjecting the aneuploid cell to an activation step;
• and
• subsequent to maintaining the aneuploid cell in the suitable environment, treating said reprogrammed aneuploid cell so as to generate a reprogrammed diploid mammalian cell from said reprogrammed aneuploid cell by removal or destruction of the mammalian recipient cell nucleus, pronucleus, metaphase plate, chromatin, chromosomes or nuclear DNA from said reprogrammed aneuploid cell;
• and
• (b) generating one of a group consisting of a cell and a cell line, from said reprogrammed diploid mammalian cell.

A method of preparing a reprogrammed mammalian cell comprising

• providing a diploid mammalian donor nucleus derived from a somatic cell, an exogenous nucleic acid molecule and a mammalian recipient cell, wherein the genetically modified diploid mammalian donor nucleus and the mammalian recipient cell are of the same species;
• introducing the mammalian donor nucleus and exogenous nucleic acid molecule into the mammalian recipient cell to produce an aneuploid cell;
• maintaining the aneuploid cell in a suitable environment for a period sufficient to allow the donor nucleus to be reprogrammed;
• subjecting the aneuploid cell to an activation step;
• and
• subsequent to maintaining the aneuploid cell in the suitable environment, treating said reprogrammed aneuploid cell so as to generate a reprogrammed diploid mammalian cell comprising an exogenous nucleic acid sequence from said reprogrammed aneuploid cell by removal or destruction of the mammalian recipient cell nucleus, pronucleus, metaphase plate, chromatin, chromosomes or nuclear DNA from said reprogrammed aneuploid cell.

A method of preparing a reprogrammed diploid embryonic mammalian cell comprising:

• providing a diploid mammalian donor nucleus derived from a somatic cell, and a mammalian recipient cell and one of a group consisting of a recipient mammalian oocyte and an embryonic cell, wherein the diploid mammalian donor nucleus and the mammalian recipient cell are of the same species;
• introducing the mammalian donor cell nucleus into the mammalian recipient oocyte or embryonic cell to produce an aneuploid cell;
• maintaining the aneuploid cell in a suitable environment for a period sufficient to allow the mammalian donor cell nucleus to be reprogrammed;
• and
• subjecting the aneuploid cell to an activation step;
• subsequent to maintaining the aneuploid cell in the suitable environment, treating said reprogrammed aneuploid cell so as to generate a reprogrammed diploid embryonic mammalian cell from said reprogrammed aneuploid cell by removal or destruction of the mammalian recipient cell nucleus, pronucleus, metaphase plate, chromatin, chromosomes or nuclear DNA from said reprogrammed aneuploid cell or one or more of its daughter cells.