Agents and methods for inhibiting human pluripotent stem cell growth

Abstract

The present invention relates to compositions and methods for inhibiting or suppressing undifferentiated or pluripotent stem cell growth and proliferation in a differentiated or differentiating cell population or culture.

What is claimed is: 
     
         1 . An in vitro composition comprising a combination of:
 a) at least one human pluripotent stem cell wherein the pluripotent cell has the ability to differentiate into cells of all three germ layers; and   b) an effective amount of a pluripotent stem cell-selective inhibitor or cytotoxic agent selected from the group consisting of caffeic acid, ivermectin, chelerythrine chloride or a mixture thereof, wherein the effective amount inhibits pluripotent cell growth in the composition as compared to pluripotent cell growth in a composition that does not comprise the effective amount of the inhibitor or cytotoxic agent.   
     
     
         2 . The composition of  claim 1 , wherein the human pluripotent stem cell is a human ES cell or a human iPS cell. 
     
     
         3 . The composition of  claim 1 , further comprising differentiating or differentiated cells, wherein the effective amount of the pluripotent stem cell-selective inhibitor or cytotoxic agent has no effect on the proliferation or viability of the differentiating or differentiated cells in the composition. 
     
     
         4 . The composition of  claim 1 , further comprising differentiating or differentiated cells, wherein the effective amount of the pluripotent stem cell-selective inhibitor or cytotoxic agent does not inhibit the differentiation or differentiation potential of the differentiating or differentiated cells in the composition. 
     
     
         5 . The composition of  claim 1 , wherein the effective amount is selected from the group consisting of at least about 1 μM caffeic acid, at least about 1 μM ivermectin or at least about 5 μM chelerythrine chloride. 
     
     
         6 . The composition of  claim 1 , further comprising differentiating or differentiated cells, wherein the differentiating or differentiated cells comprise at least one cell type selected from the group consisting of: definitive endoderm cells, PDX1-negative foregut endoderm cells, PDX1-positive foregut endoderm cells, PDX1-positive pancreatic endoderm cells, endocrine progenitor cells and endocrine precursor cells. 
     
     
         7 . An in vitro composition comprising a combination of:
 a) at least one human pluripotent stem cell, wherein the pluripotent cell has the ability to differentiate into cells of all three germ layers; and   b) an effective amount of an inhibitor of a cell signaling pathway, a phosphorylation event, or a receptor, wherein the effective amount inhibits pluripotent stem cell growth in the composition as compared to pluripotent stem cell growth in a composition that does not comprise the effective amount of the inhibitor.   
     
     
         8 . The composition of  claim 7 , wherein the human pluripotent stem cell is a human ES cell or a human iPS cell. 
     
     
         9 . The composition of  claim 7 , further comprising differentiating or differentiated cells, wherein the effective amount of the inhibitor of the cell signaling pathway, phosphorylation event, or receptor has no effect on the proliferation or viability of differentiating or differentiated cells in the composition. 
     
     
         10 . The composition of  claim 7 , further comprising differentiating or differentiated cells, wherein the effective amount of the inhibitor of the cell signaling pathway, phosphorylation event, or receptor does not inhibit the differentiation or differentiation potential of differentiating or differentiated cells in the composition. 
     
     
         11 . The composition of  claim 7 , wherein the inhibitor of the cell signaling pathway, phosphorylation event, or receptor is selected from the group consisting of caffeic acid, ivermectin, chelerythrine chloride or a mixture thereof. 
     
     
         12 . The composition of  claim 7 , wherein the effective amount is selected from the group consisting of at least about 1 μM caffeic acid, at least about 1 μM ivermectin or at least about 5 μM chelerythrine chloride. 
     
     
         13 . The composition of  claim 7 , further comprising differentiating or differentiated cells, wherein the differentiating or differentiated cells comprise at least one cell type selected from the group consisting of: definitive endoderm cells, PDX1-negative foregut endoderm cells, PDX1-positive foregut endoderm cells, PDX1-positive pancreatic endoderm cells, endocrine progenitor cells and endocrine precursor cells. 
     
     
         14 . An in vitro composition comprising a combination of:
 a) at least one human pluripotent stem cell wherein the pluripotent cell has the ability to differentiate into cells of all three germ layers; and   b) an effective amount of a pluripotent stem cell-selective inhibitor or cytotoxic agent wherein the effective amount inhibits pluripotent cell growth in the composition as compared to pluripotent cell growth in a composition that does not comprise the effective amount of the inhibitor or cytotoxic agent.   
     
     
         15 . The composition of  claim 14 , wherein the human pluripotent stem cell is a human ES cell or a human iPS cell. 
     
     
         16 . The composition of  claim 14 , further comprising differentiating or differentiated cells, wherein the effective amount of the pluripotent stem cell-selective inhibitor or cytotoxic agent has no effect on the proliferation or viability of the differentiating or differentiated cells in the composition. 
     
     
         17 . The composition of  claim 14 , further comprising differentiating or differentiated cells, wherein the effective amount of the pluripotent stem cell-selective inhibitor or cytotoxic agent does not inhibit the differentiation or differentiation potential of the differentiating or differentiated cells in the composition. 
     
     
         18 . The composition of  claim 14 , wherein the pluripotent stem cell-selective inhibitor or cytotoxic agent is selected from the group consisting of caffeic acid, ivermectin, chelerythrine chloride or a mixture thereof. 
     
     
         19 . The composition of  claim 14 , wherein the effective amount is selected from the group consisting of at least about 1 μM caffeic acid, at least about 1 μM ivermectin or at least about 5 μM chelerythrine chloride. 
     
     
         20 . The composition of  claim 14 , further comprising differentiating or differentiated cells, wherein the differentiating or differentiated cells comprise at least one cell type selected from the group consisting of: definitive endoderm cells, PDX1-negative foregut endoderm cells, PDX1-positive foregut endoderm cells, PDX1-positive pancreatic endoderm cells, endocrine progenitor cells and endocrine precursor cells.