Isolated primate embryonic cells derived from extended blastocysts
First Claim
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1. An isolated primate embryonic cell line that exhibits for at least 20 passages an undifferentiated proliferative state, the ability to differentiate to derivatives of each of an endoderm, mesoderm, and ectoderm tissue and a double staining expression of brachyury and Octamer binding transcription factor 4 (OCT-4), but not of SSEA-1.
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Abstract
An isolated primate embryonic cell is provided as well as cell cultures and cell lines derived therefrom. Also provided are methods of generating and using such cells.
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Citations
36 Claims
- 1. An isolated primate embryonic cell line that exhibits for at least 20 passages an undifferentiated proliferative state, the ability to differentiate to derivatives of each of an endoderm, mesoderm, and ectoderm tissue and a double staining expression of brachyury and Octamer binding transcription factor 4 (OCT-4), but not of SSEA-1.
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7. A method of generating a primate embryonic cell culture comprising;
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(a) providing a pre-implantation stage blastocyst; (b) ex vivo culturing said pre-implantation stage blastocyst on fibroblast feeder cells or on a synthetic extracellular matrix for at least nine days to fourteen days post fertilization to produce a pre-gastrulation, extended blastocyst; (c) isolating cells from said extended blastocyst; and (d) culturing said isolated extended blastocyst cells on fibroblast feeder cells, an extracellular matrix or media comprising basic fibroblast growth factor, thereby generating a primate embryonic cell culture comprising embryonic cells that express brachyury and Oct-4, are maintained in an undifferentiated proliferative states for at least 20 passages and have the capability to develop in to each of the three germ layers;
endoderm, mesoderm and ectoderm. - View Dependent Claims (17, 18, 19, 23, 25, 26, 32, 33)
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8. A method of generating a primate embryonic cell line comprising;
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(a) providing a pre-implantation stage blastocyst; (b) ex vivo culturing said pre-implantation stage blastocyst on fibroblast feeder cells or on a synthetic extracellular matrix for at least nine days to fourteen days post fertilization to produce an extended blastocyst (c) isolating cells from said extended blastocyst; (d) culturing said isolated extended blastocyst cells on fibroblast feeder cells, an extracellular matrix or media comprising basic fibroblast growth factor; and (d) cloning at least one of said isolated cells, thereby generating a primate embryonic cell line comprising embryonic cells that express brachyury and Oct-4, are maintained in an undifferentiated proliferative state for at least 20 passages and have the capability to develop in to each of the three germ layers;
endoderm, mesoderm and ectoderm. - View Dependent Claims (24, 27, 28, 34, 35)
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- 29. An isolated clonal primate embryonic cell line comprising cells that exhibit for at least 20 passages an undifferentiated proliferative state, the ability to differentiate to derivatives of each of an endoderm, mesoderm, and ectoderm tissue and a double staining expression of brachyury and Octamer binding transcription factor 4 (OCT-4), but not of SSEA-1.
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30. An isolated primate embryonic cell line that exhibits for at least 20 passages an undifferentiated proliferative state, the ability to differentiate to derivatives of each of an endoderm, mesoderm, and ectoderm tissue and a double staining expression of brachyury and Octamer binding transcription factor 4 (OCT-4), but not of SSEA-1, wherein the cell line is not genetically modified.
Specification