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, CD90, CD66A, CD66E, CXCR4, CD133 or an SSEA. The invention further provides a proliferative stem cell, which expresses genes including Oct-4, Nanog, Sox2, GDF3, P16INK4, BMI, Notch, HDAC4, TERT, Rex-1, TWIST, KLF-4 and Stella but does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CF34, CF49c, CD73, CD105, CD90, CD66A, CD66E, CXCR4, CD133 or an SSEA. 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 stem cells, are also provided.

1 . An isolated adult stem cell that is capable of proliferating and differentiating into at least two of ectoderm, mesoderm, or endoderm, expresses at least one of Oct-4, KLF-4, Nanog, Sox-2, Rex-1, GDF-3, and Stella, and does not detectibly express CD13, CD45, CD90, and CD34 and further does not detectibly express at least one of MHC class I, MHC class II, CD44, CD105, CD49c, CD73, CD66A, CD66E, CXCR4, CD133 or an SSEA. 
     
     
         2 . An isolated quiescent adult stem cell that is capable of proliferating and differentiating into at least two of ectoderm, mesoderm, and endoderm and does not detectibly express Oct-4, CD13, CD45, CD90, CD34 and further does not detectibly express at least one of MHC class I, MHC class II, CD44, CD105, CD49c, CD73, CD66A, CD66E, CXCR4, CD133 or an SSEA. 
     
     
         3 . The cell of  claim 1 , wherein the SSEA is SSEA-4. 
     
     
         4 . The cell of  claim 1 , wherein said cell is synovial fluid derived, blood derived or tissue derived. 
     
     
         5 . The cell of  claim 1 , wherein said cell is isolated from a mammal. 
     
     
         6 . A population of isolated adult stem cells that are capable of proliferating and differentiating into at least two of ectoderm, mesoderm, and endoderm, express at least one of Oct-4, KLF-4, Nanog, Sox-2, Rex-1, GDF-3, and Stella, and does not detectibly express CD13, CD45, CD90, CD34 and further does not detectibly express at least one of MHC class I, MHC class II, CD44, CD105, CD49c, CD73, CD66A, CD66E, CXCR4, CD133 or an SSEA, wherein from about 10% to about 30% of the population of adult stem cells are quiescent. 
     
     
         7 . The population of  claim 6 , wherein said population is a culture expanded population. 
     
     
         8 . The population of  claim 6 , wherein said population comprises said stem cells in an amount of at least 1×103, at least 1×106, at least 1×109, at least 1×1012, or at least 1×1014. 
     
     
         9 . The population of  claim 6 , further comprising a bioactive compound. 
     
     
         10 . A master cell bank comprising a plurality of cryopreserved individually packaged populations of isolated adult stem cells, each population including at least 1×102 or more of the cells of  claim 1 . 
     
     
         11 . A method of forming an adipocyte, the method comprising culturing a stem cell of  claim 1  under adipocyte-differentiating conditions. 
     
     
         12 . The method of  claim 11 , wherein said adipocyte-differentiating conditions include culturing with at least one of dexamethasone, 3-isobutyl-1-methylxanthine (IBMX), insulin, and indomethacin. 
     
     
         13 . A method of forming a muscle cell, said method comprising culturing a stem cell of  claim 1  under muscle cell differentiating conditions. 
     
     
         14 . A method of forming a neural cell, said method comprising the steps of contacting a stem cell of  claim 1  under neural cell-forming conditions. 
     
     
         15 . A method of forming a hepatocyte, said method comprising culturing a stem cell of  claim 1  under hepatocyte-forming conditions. 
     
     
         16 . A method of forming an endothelial cell, said method comprising culturing a stem cell of  claim 1  under endothelial cell-forming conditions. 
     
     
         17 . A method of forming a hematopoietic cell, the method comprising culturing a stem cell of  claim 1  under hematopoietic cell forming conditions. 
     
     
         18 . The method of  claim 17 , wherein said conditions include culturing said cell with bone morphogenic protein-4 (BMP4), VEGF, bFGF, stem cell factor (SCF), F1t3L, hyper IL6, thrombopoietin (TPO) and erythropoietin (EPO). 
     
     
         19 . 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. 
     
     
         20 . The method of  claim 19 , wherein said administration of said cells results in reduced scarring at the wound site. 
     
     
         21 . A process for expanding the population of  claim 6  comprising passaging the population at least three, at least five, at least ten or at least forty times. 
     
     
         22 . A population of stern cells obtained according to the process of  claim 21 .