Isolated homozygous stem cells, differentiated cells derived therefrom, and materials and methods for making and using same
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Abstract
The present invention discloses and describes pluripotent homozygous stem (HS) cells, and methods and materials for making same. The present invention also provides methods for differentiation of HS cells into progenitor (multipotent) cells or other desired cells, groups of cells or tissues. Further, the applications of the HS cells disclosed herein, include (but are not limited to) the diagnosis and treatment of various diseases (for example, genetic diseases, neurodegenerative diseases, endocrine-related disorders and cancer), traumatic injuries, cosmetic or therapeutic transplantation, gene therapy and cell replacement therapy.
91 Citations
72 Claims
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1. An isolated homozygous stem cell (HS).
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2. An isolated homozygous stem cell (HS) derived from human.
- 3. An isolated homozygous stem cell (HS) derived from non-human species.
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5. An isolated homozygous stem cell derived from a method comprising:
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(a) producing a mitotically activated homozygous post-meiosis I diploid germ cell by;
fusing two oocytes or two spermatids, preventing the extrusion of the second polar body during oogenesis, allowing the extrusion of the second polar body and spontaneous self-replication under appropriate conditions, or transferring two sperm or two haploid egg nuclei into an enucleated oocyte;
(b) culturing said activated homozygous post-meiosis I diploid germ cell to form a blastocyst-like mass; and
,(c) isolating homozygous stem cells from the inner cell mass of said blastocyst-like mass. - View Dependent Claims (6)
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7. A method of producing homozygous stem cells comprising:
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(a) producing a mitotically activated homozygous post-meiosis I diploid germ cell by;
fusing two oocytes or two spermatids, preventing the extrusion of the second polar body during oogenesis, allowing the extrusion of the second polar body and spontaneous self-replication under appropriate conditions, or transferring two sperm or two haploid egg nuclei into an enucleated oocyte;
(b) culturing said activated homozygous post-meiosis I diploid germ cell to form a blastocyst-like mass; and
,(c) isolating homozygous stem cells from the inner cell mass of said blastocyst-like mass. - View Dependent Claims (8)
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- 9. A method of making a desired progenitor cell, differentiated cell, group of differentiated cells, or tissue type comprising inducing isolated homozygous stem cells to differentiate under suitable conditions.
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33. A method of producing progenitor cells, comprising:
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(a) producing a mitotically activated homozygous post-meiosis I diploid germ cell by;
fusing two oocytes or two spermatids, preventing the extrusion of the second polar body during oogenesis, allowing the extrusion of the second polar body and spontaneous self-replication under appropriate conditions, or transferring two sperm or two haploid egg nuclei into an enucleated oocyte;
(b) culturing said activated homozygous post-meiosis I diploid germ cell to form a blastocyst-like mass;
(c) isolating homozygous stem cells from the inner cell mass of said blastocyst-like mass; and
,(d) inducing differentiation of said homozygous stem cells to produce progenitor cells. - View Dependent Claims (34, 35, 36, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52)
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37. A method for producing genetically altered progenitor cells, comprising:
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(a) inserting, removing or modifying a desired gene in an HS cell to create a genetically altered HS cell; and
,(b) inducing differentiation of said genetically altered HS cell.
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38. A method for producing genetically altered progenitor cells, comprising:
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(a) inserting, removing or modifying a desired gene in a progenitor cell derived from an HS cell to create a genetically altered progenitor cell; and
(b) culturing said genetically altered progenitor cell to grow genetically altered progenitor cells.
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53. A progenitor cell derived from a method comprising:
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(a) producing a mitotically activated homozygous post-meiosis I diploid germ cell;
(b) culturing said activated homozygous post-meiosis I diploid germ cell to form a blastocyst-like mass;
(c) isolating homozygous stem cells from the inner cell mass of said blastocyst-like mass; and
,(d) inducing differentiation of said homozygous stem cells to produce progenitor cells. - View Dependent Claims (54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69)
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70. An isolated homozygous stem cell derived from a method comprising:
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(a) producing a mitotically activated homozygous post-meiosis I diploid germ cell by;
fusing two oocytes or two spermatids, preventing the extrusion of the second polar body during oogenesis, allowing the extrusion of the second polar body and spontaneous self-replication under appropriate conditions, or transferring two sperm or two haploid egg nuclei into an enucleated oocyte;
(b) culturing said activated homozygous post-meiosis I diploid germ cell to form a blastocyst-like mass being surrounded by zona pellucida;
(c) releasing said blastocyst-like mass from said zona pellucida using the method of assisted hatching; and
,(d) isolating homozygous stem cells from the inner cell mass of said blastocyst-like mass. - View Dependent Claims (71)
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72. An isolated homozygous stem cell derived from a method comprising:
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(a) producing a mitotically activated homozygous post-meiosis I diploid germ cell by;
fusing two oocytes or two spermatids, preventing the extrusion of the second polar body during oogenesis, allowing the extrusion of the second polar body and spontaneous self-replication under appropriate conditions, or transferring two sperm or two haploid egg nuclei into an enucleated oocyte;
(b) culturing said activated homozygous post-meiosis I diploid germ cell to form a blastocyst-like mass being surrounded by zona pellucida;
(c) transferring said activated post-meiosis I diploid germ cell on day 2 post-activation to a mitomycin C treated feeder layer of mouse embryonic fibroblasts until a blastocyst-like mass is formed;
(d) releasing said blastocyst-like mass from said zona pellucida using the method of assisted hatching; and
,(e) isolating homozygous stem cells from the inner cell mass of said blastocyst-like mass.
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Specification