Multipotent stem cells and uses thereof

Abstract

The invention provides a quiescent stem cell having the capacity to differentiate into ectoderm, mesoderm and endoderm, and which does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105 and CD90. The invention further provides a proliferative stem cell, which expresses genes including Oct-4, Nanog, Sox2, GDF3, P16INK4, BMI, Notch, HDAC4, TERT, Rex-1 and TWIST but does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105 and CD90. The cells of the invention can be isolated from adult mammals, have embryonic cell characteristics, and can form embryoid bodies. Methods for obtaining the stem cells, as well as methods of treating diseases and differentiated the stem cells, are also provided.

1 . An isolated stem cell which is capable of proliferating and differentiating into ectoderm, mesoderm, and endoderm, expresses at least one of Oct-4, Nanog, Sox-2, Rex-1, GDF-3, and Stella and does not express CD13, CD44, CD45, CD90, and CD105. 
     
     
         2 . An isolated quiescent stem cell which is capable of proliferating and differentiating into ectoderm, mesoderm, and endoderm and does not express Oct-4, CD13, CD44, CD45, CD90, and CD105. 
     
     
         3 . The cell of  claim 1 , wherein said cell further does not express at least one of MHC class I, MHC class II, CD34, CD49c, and CD73. 
     
     
         4 . The cell of  claim 1 , wherein said cell is isolated from synovial fluid, blood or tissue. 
     
     
         5 . The cell of  claim 1 , wherein said cell is substantially purified. 
     
     
         6 . The cell of  claim 1 , wherein said cell is isolated from a mammal. 
     
     
         7 . The cell of  claim 6 , wherein said cell is isolated from a human. 
     
     
         8 . The cell of  claim 1 , wherein said cell is isolated from an adult mammal. 
     
     
         9 . The cell of  claim 1 , comprising a heterologous nucleic acid sequence. 
     
     
         10 . The cell of  claim 9 , wherein said heterologous nucleic acid sequence comprises a stem cell-specific promoter. 
     
     
         11 . The cell of  claim 10 , wherein said promoter is the promoter of an embryonic transcription factor. 
     
     
         12 . The cell of  claim 11 , wherein said promoter is selected from the group consisting of the Oct-4 promoter, Nanog promoter, Sox2 promoter, cMyc-2 promoter, KLF4 promoter, Rex-1 promoter, GDF-3 promoter, Stella promoter, FoxD3 promoter, Polycomb Repressor Complex 2 promoter, and CTCF promoter. 
     
     
         13 . The cell of  claim 10 , wherein said promoter is operably linked to a detectable gene product. 
     
     
         14 . The cell of  claim 13 , wherein said detectable gene product is GFP. 
     
     
         15 . A population of cells wherein at least 10% of the cells are the stem cells of  claim 1 . 
     
     
         16 . (canceled) 
     
     
         17 . A composition comprising:
 (a) a population of cells of  claim 15 ; and   (b) a cryoprotectant, a dendritic cell or an antigen presenting cell or a committed or differentiated cell.   
     
     
         18 - 20 . (canceled) 
     
     
         21 . A method of isolating a population of stem cells, said method comprising the steps:
 (a) providing a bodily fluid or tissue from a subject;   (b) enriching for a population of cells that are about 6 μm to 20 μm in size; and   (c) depleting cells from said population expressing a stem cell surface marker or an MHC protein, thereby isolating a population of stem cells.   
     
     
         22 - 37 . (canceled) 
     
     
         38 . A method of forming a muscle cell, said method comprising culturing a stem cell of  claim 1  under muscle cell differentiating conditions. 
     
     
         39 . (canceled) 
     
     
         40 . A method of forming a neural cell, said method comprising culturing a stem cell of  claim 1  under neural cell-forming conditions. 
     
     
         41 . (canceled) 
     
     
         42 . A method of forming a hematopoietic cell, the method comprising culturing a stem cell of  claim 1  under hematopoietic cell-forming conditions. 
     
     
         43 . (canceled) 
     
     
         44 . A method of forming an endothelial cell, the method comprising culturing a stem cell of  claim 1  under endothelial cell-forming conditions. 
     
     
         45 . (canceled) 
     
     
         46 . A method of forming a hematopoietic cell, the method comprising culturing a stem cell of  claim 1  under hematopoietic cell-forming conditions. 
     
     
         47 . (canceled) 
     
     
         48 . (canceled) 
     
     
         49 . A method for promoting wound healing in a subject, said method comprising administering a stem cell of  claim 1 , or a committed or differentiated progeny thereof, to said wound or to a site near said wound in an amount sufficient to promote the healing of said wound. 
     
     
         50 . (canceled) 
     
     
         51 . A method for treating a cardiovascular disease in a subject, said method comprising administering to said subject a stem cell of  claim 1 , or a committed or differentiated progeny thereof, in an amount sufficient to treat said disease. 
     
     
         52 . (canceled) 
     
     
         53 . A method of increasing vascularization in a subject, said method comprising administering to said subject a stem cell of  claim 1 , or a committed or differentiated progeny thereof, in an amount sufficient to increase vascularization. 
     
     
         54 . (canceled) 
     
     
         55 . A method for treating a neurological disorder in a subject, said method comprising administering to said subject a stem cell of  claim 1 , or a committed or differentiated progeny thereof, in an amount sufficient to treat said disease. 
     
     
         56 . (canceled) 
     
     
         57 . (canceled) 
     
     
         58 . A method for treating an autoimmune disease in a subject, said method comprising administering to said subject a stem cell of  claim 1 , or a committed or differentiated progeny thereof, in an amount sufficient to treat said disease. 
     
     
         59 . A method for reducing or preventing rejection of transplanted tissue in a subject, said method comprising administering to said subject a stem cell of  claim 1  or a committed or differentiated progency thereof, in an amount sufficient to reduce or prevent said rejection.