MULTIPOTENT/PLURIPOTENT CELLS AND METHODS
First Claim
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1. A human stem cell that is pluripotent, somatic, non-embryonic, and having the property of long-term self renewal.
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Abstract
Described herein are multipotent stem cells, e.g., human and other mammalian pluripotent stem cells, and related methods.
122 Citations
159 Claims
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1. A human stem cell that is pluripotent, somatic, non-embryonic, and having the property of long-term self renewal.
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2-132. -132. (canceled)
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133. A method of creating a clonal culture of human pluripotent stem cells, comprising the steps of:
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providing a culture of human pluripotent stem cells in culture in a culture medium in a culture vessel; selecting a single cell from the stem cells in culture; and initiating a new culture of the stem cells in a culture vessel containing a matrix with the single cell in a defined culture medium capable of supporting growth of pluripotent cells, the culture medium including an agent selected to increase the cloning efficiency of the stem cell culture relative to a stem cell culture that does not contain the agent, wherein the agent is a small molecule and wherein the agent is a kinase inhibitor. - View Dependent Claims (134, 135, 136)
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137. A method of creating a clonal culture of human pluripotent stem cells, comprising the steps of:
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providing a culture of human pluripotent stem cells in culture in a defined medium capable of supporting growth of pluripotent cells in a culture vessel; selecting a single cell from the stem cells in culture; and initiating a new culture of the human pluripotent stem cells in the defined medium in a culture vessel containing a matrix with the single cell, the culture medium including an agent selected to increase the cloning efficiency of the stem cell culture relative to a stem cell culture that does not contain the agent, the agent being selected from the group consisting of - View Dependent Claims (138)
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139. A human pluripotent stem cell culture, comprising:
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a culture vessel containing a matrix; a defined culture medium capable of supporting growth of pluripotent cells contained in the vessel; human pluripotent stem cells growing in the culture medium; and the culture medium containing an agent that is a kinase inhibitor. - View Dependent Claims (140, 141)
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- 142. An improvement in a medium in which to culture human pluripotent stem cells, the improvement comprising including in the medium an agent selected to increase the cloning efficiency of the stem cell culture, wherein the agent is selected from the group consisting of
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143. A method of culturing human pluripotent stem cells, comprising the steps of:
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(a) culturing human pluripotent stem cells in a culture medium in a culture vessel; (b) selecting a single cell from the human pluripotent stem cells in culture; and (c) initiating a new culture of the single cell in a culture vessel, wherein the culture vessel contains a matrix and a culture medium capable of supporting growth of human pluripotent cells, and wherein the culture medium includes an agent that is a kinase inhibitor.
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147. A method of culturing human pluripotent stem cells, comprising the steps of:
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(a) culturing human pluripotent stem cells in a defined medium capable of supporting growth of pluripotent cells in a culture vessel; (b) selecting a single cell from the stem cells in culture; and (c) initiating a new culture of the single cell in a culture vessel, wherein the culture vessel contains a matrix and a culture medium capable of supporting growth of human pluripotent cells, and wherein the culture medium includes an agent that is selected from the group consisting of HA-1077 and Y-27632.
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149. A human pluripotent stem cell culture comprising:
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(a) a culture vessel containing a matrix; (b) a culture medium capable of supporting growth of pluripotent cells contained in the vessel; (c) human pluripotent stem cells growing in the culture medium; and (d) the culture medium contains a kinase inhibitor.
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152. An improvement in a medium in which to culture human pluripotent stem cells, the improvement comprising including in the medium an agent selected to increase the cloning efficiency of the stem cell culture, wherein the agent is selected from the group consisting of HA-1077 and Y-27632.
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153. A method of culturing human pluripotent stem cells, comprising the steps of:
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(a) culturing pluripotent stem cells in a culture medium in a culture vessel; (b) selecting a single cell from the pluripotent stem cells in culture; and (c) initiating a new culture of the single cell in a culture vessel containing a substrate, wherein the substrate is coated with fibronectin, gelatin, matrigel™
(BD Bioscience), collagen, or laminin, wherein the single cell is cultured in a culture medium that is capable of supporting growth of pluripotent cells, and wherein the culture medium includes a kinase inhibitor. - View Dependent Claims (154, 155, 156, 157, 158, 159)
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Specification