Method of cloning bovines using reprogrammed non-embryonic bovine cells
AssigneeInfigen, Inc.
InventorsNikolai S. Strelchenko, Jeffrey M. Betthauser, Gail L. Jurgella, Marvin M. Pace, Michael D. Bishop
Life of this patent
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Abstract
The present invention relates to cloning technologies. The invention relates in part to immortalized and totipotent cells useful for cloning animals, the embryos produced from these cells using nuclear transfer techniques, animals that arise from these cells and embryos, and materials, methods, and processes for establishing such cells, embryos, and animals.
Claims29
A method of cloning a bovine, said method comprising:
- a) obtaining a cell from a bovine fetus of not less than about 35 days of gestation and not more than about 70 days of gestation;
- b) reprogramming said cell by incubating said cell in a medium comprising LIF and FGF to form a reprogrammed cell;
- c) forming a cybrid by nuclear transfer of said reprogrammed cell into an enucleated bovine oocyte;
- and
- d) culturing said cybrid so as to generate an embryo comprising embryonic cells;
- and
- e) transferring said embryo of step (d) or a recloned embryo of said embryo of step (d) into the uterus of a host bovine so as to produce a fetus which undergoes full fetal development and parturition;
- whereby a viable clone of said bovine is generated.
A method of producing a totipotent bovine cell line comprising:
- a) obtaining non-embryonic bovine cells;
- b) reprogramming said cells by incubating said cells in a medium comprising LIF and FGF to form reprogrammed cells;
- and
- c) culturing said reprogrammed cells as a totipotent cell line.
A method of producing a totipotent bovine cell line comprising:
- a) obtaining non-embryonic bovine cells;
- b) reprogramming said cells by incubating said cells in a medium comprising LIE and FGF to form reprogranmed cells;
- c) fusing a said reprogrammed cell with an enucleated young oocyte by performing a nuclear transfer to form a cybrid;
- d) culturing said cybrid so as to generate an embryo comprising embryonic cells;
- and
- e) culturing said embryonic cells to generate a totipotent cell line.
A method for cloning a bovine, comprising the steps of:
- a) obtaining a non-embryonic bovine cell;
- b) reprogramming said non-embryonic bovine cell cells by incubating said cell in a medium comprising LIF and FGF to form a reprogrammed cell, wherein said reprogrammed cell is immortalized and totipotent;
- c) forming a cybrid by nuclear transfer of said reprogrammed cell into an enucleated bovine oocyte;
- and
- d) culturing said cybrid so as to generate an embryo comprising embryonic cells;
- and
- e) transferring said embryo of step (d) or a recloned embryo of said embryo of step (d) into the uterus of a host bovine for developing said bovine.
Dependencies are inferred from the claim text.