Method of dynamically culturing embryonic stem cells
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Abstract
he present invention is of a method of dynamically generating human embryoid bodies which can be used for generating lineage specific cells and cell lines. Specifically, the present invention can be used to generate ESC-differentiated cells for cell-replacement therapy.
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Citations
109 Claims
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1-76. -76. (canceled)
- 77. A method of dynamically generating embryoid bodies comprising culturing embryonic stem cells in a bioreactor selected capable of subjecting cells cultured in a fluid contained therein to randomized gravity vectors without formation of gas bubbles in said fluid to thereby obtain dynamically generated embryoid bodies.
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78. A method of generating expanded and/or differentiated cells from embryonic stem cells comprising:
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(a) culturing embryonic stem cells in a bioreactor selected capable of subjecting cells cultured in a fluid contained therein to randomized gravity vectors without formation of gas bubbles in said fluid to thereby obtain dynamically generated embryoid bodies;
(b) isolating lineage specific cells from said embryoid bodies, and;
(c) culturing said lineage specific cells under culturing conditions selected suitable for the expansion and/or differentiation of said lineage specific cells to thereby obtain expanded and/or differentiated lineage-specific cells.
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- 93. A cell culture comprising embryoid bodies, wherein at least 90% of said embryoid bodies fall within a diameter size range of 400-800 μ
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95. A method of treating a disorder requiring cell replacement therapy comprising:
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(a) culturing embryonic stem cells in a bioreactor selected capable of subjecting cells cultured in a fluid contained therein to randomized gravity vectors without formation of gas bubbles in said fluid to thereby obtain dynamically generated embryoid bodies;
(b) isolating lineage specific cells from said embryoid bodies;
(c) culturing said lineage specific cells under culturing conditions selected suitable for the expansion and/or differentiation of said lineage specific cells to thereby obtain expanded and/or differentiated lineage-specific cells, and;
(d) administering cells of said expanded and/or differentiated lineage-specific cells to an individual in need thereof thereby treating the disorder requiring cell replacement therapy. - View Dependent Claims (96, 97, 98, 99, 100, 101, 102)
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- 103. A method of dynamically expanding embryonic stem cells while maintaining the embryonic stem cells in an undifferentiated state, the method comprising culturing embryonic stem cells in a bioreactor selected capable of subjecting cells cultured in a fluid contained therein to randomized gravity vectors without formation of gas bubbles in said fluid to thereby obtain dynamically expanded undifferentiated embryonic stem cell cultures.
Specification