Nuclear reprogramming factor and induced pluripotent stem cells

Abstract

The present invention relates to a nuclear reprogramming factor having an action of reprogramming a differentiated somatic cell to derive an induced pluripotent stem (iPS) cell. The present invention also relates to the aforementioned iPS cells, methods of generating and maintaining iPS cells, and methods of using iPS cells, including screening and testing methods as well as methods of stem cell therapy. The present invention also relates to somatic cells derived by inducing differentiation of the aforementioned iPS cells.

1 . A pluripotent stem cell induced by reprogramming a somatic cell, wherein the reprogramming is performed in the absence of eggs, embryos, or embryonic stem (ES) cells and comprises contacting the somatic cell with a nuclear reprogramming factor comprising one or more gene products of a Sall1 gene and/or a Sall4 gene. 
     
     
         2 . The pluripotent stem cell of  claim 1 , wherein the somatic cell is a human cell. 
     
     
         3 . The pluripotent stem cell of  claim 1 , wherein the somatic cell is a mouse cell. 
     
     
         4 . The pluripotent stem cell of  claim 1 , wherein the nuclear reprogramming factor is encoded by a gene contained in a recombinant vector introduced into the cell. 
     
     
         5 . The pluripotent stem cell of  claim 1 , wherein the nuclear reprogramming factor comprises one or more gene products of a human or mouse Sall4 gene. 
     
     
         6 . The pluripotent stem cell of  claim 1 , wherein the nuclear reprogramming factor comprises one or more gene products of each of: an Oct family gene, a Klf family gene, and a Sox family gene. 
     
     
         7 . The pluripotent stem cell of  claim 6 , wherein the nuclear reprogramming factor comprises one or more gene products of a Myc family gene. 
     
     
         8 . The pluripotent stem cell of  claim 6 , wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, and Sox2, and at least one of Sall1 and Sall4. 
     
     
         9 . The pluripotent stem cell of  claim 7 , wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, Sox2, and c-Myc, and at least one of Sall1 and Sall4. 
     
     
         10 . The pluripotent stem cell of  claim 1 , wherein the nuclear reprogramming factor comprises one or more gene products of each of: an Oct family gene, a Sox family gene, and a Nanog family gene. 
     
     
         11 . The pluripotent stem cell of  claim 10 , wherein the nuclear reprogramming factor includes one or more gene products of a Lin28 family gene. 
     
     
         12 . The pluripotent stem cell of  claim 10 , wherein the nuclear reprogramming factor comprises Oct3/4, Sox2, and Nanog, and at least one of Sall1 and Sall4. 
     
     
         13 . The pluripotent stem cell of  claim 11 , wherein the nuclear reprogramming factor comprises Oct3/4, Sox2, Nanog, and Lin28, and at least one of Sall1 and Sall4. 
     
     
         14 . A pluripotent stem cell induced by reprogramming a somatic cell, wherein the reprogramming is performed in the absence of eggs, embryos, or embryonic stem (ES) cells and comprises contacting the somatic cell with a nuclear reprogramming factor comprising one or more gene products of: an L-Myc gene, an N-Myc gene, and/or a c-Myc gene. 
     
     
         15 . The pluripotent stem cell of  claim 14 , wherein the somatic cell is a human cell. 
     
     
         16 . The pluripotent stem cell of  claim 14 , wherein the somatic cell is a mouse cell. 
     
     
         17 . The pluripotent stem cell of  claim 14 , wherein the nuclear reprogramming factor is encoded by a gene contained in a recombinant vector introduced into the cell. 
     
     
         18 . The pluripotent stem cell of  claim 14 , wherein the L-Myc gene, the N-Myc gene, and/or the c-Myc gene is a chimera or mutant gene. 
     
     
         19 . The pluripotent stem cell of  claim 18 , wherein the nuclear reprogramming factor comprises Ms-cL-Myc or its human homolog. 
     
     
         20 . The pluripotent stem cell of  claim 14 , wherein the nuclear reprogramming factor comprises amino acids 1-144 of c-Myc or its human homolog. 
     
     
         21 . The pluripotent stem cell of  claim 14 , wherein the nuclear reprogramming factor comprises amino acids 105-277 of L-Myc or its human homolog. 
     
     
         22 . The pluripotent stem cell of  claim 14 , wherein the nuclear reprogramming factor comprises amino acids 347-439 of c-Myc, amino acids 278-368 of L-Myc, or their human homolog. 
     
     
         23 . The pluripotent stem cell of  claim 14 , wherein the nuclear reprogramming factor comprises one or more gene products of: an Oct family gene, and a Klf family gene. 
     
     
         24 . The pluripotent stem cell of  claim 14 , wherein the nuclear reprogramming factor comprises one or more gene products of a Sox family gene. 
     
     
         25 . The pluripotent stem cell of  claim 14 , wherein the nuclear reprogramming factor comprises one or more gene products of a Lin28 family gene. 
     
     
         26 . The pluripotent stem cell of  claim 23 , wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, and L-Myc. 
     
     
         27 . The pluripotent stem cell of  claim 24 , wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, Sox2, and L-Myc. 
     
     
         28 . The pluripotent stem cell of  claim 25 , wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, Sox2, L-Myc, and Lin28. 
     
     
         29 . A pluripotent stem cell induced by reprogramming a somatic cell, wherein the reprogramming is performed in the absence of eggs, embryos, or embryonic stem (ES) cells and comprises contacting the somatic cell with a nuclear reprogramming factor comprising one or more gene products of a Lin28 gene. 
     
     
         30 . The pluripotent stem cell of  claim 29 , wherein the nuclear reprogramming factor is encoded by a gene contained in a recombinant vector introduced into the cell. 
     
     
         31 . A method for preparing an induced pluripotent stem cell by nuclear reprogramming of a somatic cell, which comprises contacting a nuclear reprogramming factor comprising one or more gene products of a Sall1 gene and/or a Sall4 gene with the somatic cell to obtain an induced pluipotent stem cell. 
     
     
         32 . The method of  claim 31 , wherein the somatic cell is a human cell. 
     
     
         33 . The method of  claim 31 , wherein the somatic cell is a mouse cell. 
     
     
         34 . The method of  claim 31 , wherein the nuclear reprogramming factor is encoded by a gene contained in a recombinant vector introduced into the cell. 
     
     
         35 . The method of  claim 31 , wherein the nuclear reprogramming factor comprises one or more gene products of a human or mouse Sall4 gene. 
     
     
         36 . The method of  claim 31 , wherein the nuclear reprogramming factor comprises one or more gene products of each of: an Oct family gene, a Klf family gene, and a Sox family gene. 
     
     
         37 . The method of  claim 36 , wherein the nuclear reprogramming factor comprises one or more gene products of a Myc family gene. 
     
     
         38 . The method of  claim 36 , wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, and Sox2, and at least one of Sall1 and Sall4. 
     
     
         39 . The method of  claim 37 , wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, Sox2, and c-Myc, and at least one of Sall1 and Sall4. 
     
     
         40 . The method of  claim 31 , wherein the nuclear reprogramming factor comprises one or more gene products of each of: an Oct family gene, a Sox family gene, and a Nanog family gene. 
     
     
         41 . The method of  claim 40 , the efficiency of the iPS cell preparation is increased by the addition to the nuclear reprogramming factor of one or more gene products of a Lin28 gene. 
     
     
         42 . The method of  claim 40 , wherein the nuclear reprogramming factor comprises Oct3/4, Sox2, and Nanog, and at least one of Sall1 and Sall4. 
     
     
         43 . The method of  claim 41 , wherein the nuclear reprogramming factor comprises Oct3/4, Sox2, Nanog, and Lin28, and at least one of Sall1 and Sall4. 
     
     
         44 . A method for preparing an induced pluripotent stem cell by nuclear reprogramming of a somatic cell, which comprises contacting a nuclear reprogramming factor comprising one or more gene products of: an L-Myc gene, an N-Myc gene, and/or a c-Myc gene with the somatic cell to obtain an induced pluipotent stem cell. 
     
     
         45 . The method of  claim 44 , wherein the somatic cell is a human cell. 
     
     
         46 . The method of  claim 44 , wherein the somatic cell is a mouse cell. 
     
     
         47 . The method of  claim 44 , wherein the nuclear reprogramming factor is encoded by a gene contained in a recombinant vector introduced into the cell. 
     
     
         48 . The method of  claim 44 , wherein the L-Myc gene, the N-Myc gene, and/or the c-Myc gene is a chimera or mutant gene. 
     
     
         49 . The method of  claim 48 , wherein the nuclear reprogramming factor comprises Ms-cL-Myc or its human homolog. 
     
     
         50 . The method of  claim 44 , wherein the nuclear reprogramming factor comprises amino acids 1-144 of c-Myc or its human homolog. 
     
     
         51 . The method of  claim 44 , wherein the nuclear reprogramming factor comprises amino acids 105-277 of L-Myc or its human homolog. 
     
     
         52 . The method of  claim 44 , wherein the nuclear reprogramming factor comprises amino acids 347-439 of c-Myc, amino acids 278-368 of L-Myc, or their human homolog. 
     
     
         53 . The method of  claim 44 , wherein the nuclear reprogramming factor comprises one or more gene products of: an Oct family gene, and a Klf family gene. 
     
     
         54 . The method of  claim 44 , wherein the nuclear reprogramming factor comprises one or more gene products of a Sox family gene. 
     
     
         55 . The method of  claim 44 , wherein the nuclear reprogramming factor comprises one or more gene products of a Lin28 family gene. 
     
     
         56 . The method of  claim 53 , wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, and L-Myc. 
     
     
         57 . The method of  claim 54 , wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, Sox2, and L-Myc. 
     
     
         58 . The method of  claim 55 , wherein the nuclear reprogramming factor comprises Oct3/4, Klf4, Sox2, L-Myc, and Lin28. 
     
     
         59 . A method for preparing an induced pluripotent stem cell by nuclear reprogramming of a somatic cell, which comprises contacting a nuclear reprogramming factor comprising one or more gene products of a Lin28 gene with the somatic cell to obtain an induced pluipotent stem cell. 
     
     
         60 . The method of  claim 59 , wherein the nuclear reprogramming factor is encoded by a gene contained in a recombinant vector introduced into the cell. 
     
     
         61 . An induced pluripotent stem cell obtained by the method of  claim 31 . 
     
     
         62 . An induced pluripotent stem cell obtained by the method of  claim 44 . 
     
     
         63 . An induced pluripotent stem cell obtained by the method of  claim 59 . 
     
     
         64 . A somatic cell derived by inducing differentiation of the pluripotent stem cell according to  claim 61 . 
     
     
         65 . A somatic cell derived by inducing differentiation of the pluripotent stem cell according to  claim 62 . 
     
     
         66 . A somatic cell derived by inducing differentiation of the pluripotent stem cell according to  claim 63 . 
     
     
         67 . A somatic cell derived by inducing differentiation of the pluripotent stem cell according to  claim 1 . 
     
     
         68 . A somatic cell derived by inducing differentiation of the pluripotent stem cell according to  claim 14 . 
     
     
         69 . A somatic cell derived by inducing differentiation of the pluripotent stem cell according to  claim 29 .