Methods of reprogramming animal somatic cells

Abstract

This invention generally relates to methods to obtain mammalian cells and tissues with patterns of gene expression similar to that of a developing mammalian embryo or fetus, and the use of such cells and tissues in the treatment of human disease and age-related conditions. More particularly, the invention relates to methods for identifying, expanding in culture, and formulating mammalian pluripotent stem cells and differentiated cells that differ from cells in the adult human in their pattern of gene expression, and therefore offer unique characteristics that provide novel therapeutic strategies in the treatment of degenerative disease.

1 - 51 . (canceled) 
     
     
         52 . A composition comprising two, three, four, or five reprogramming factors selected from the group consisting of BARX1, CROC4, DNMT3B, H2AFX, HHEX, HISTIH2AB, HISTIH4J, HMGB2, hsa-miR-18a, hsa-miR-18b, hsa-miR-20b, hsa-miR-96, hsa-miR-106a, hsa-miR-107, hsa-miR-141, hsa-miR-183, hsa-miR-187, hsa-miR-203, hsa-miR-211, hsa-miR-217, hsa-miR-218-1, hsa-miR-218-2, hsa-miR-302a, hsa-miR-302c, hsa-miR-302d, hsa-miR-330, hsa-miR-363, hsa-miR-367, hsa-miR-371, hsa-miR-372, hsa-miR-373, hsa-miR-496, hsa-miR-50B, hsa-miR-512-3p, hsa-miR-512-5p, hsa-miR-515-3p, hsa-miR-515-5p, hsa-miR-516-5p, hsa-miR-517, hsa-miR-517a, hsa-miR-518b, hsa-miR-51Bc, hsa-miR-518e, hsa-miR-51 ge, hsa-miR-520a, hsa-miR-520b, hsa-miR-520e, hsa-miR-520 g, hsa-miR-520h, hsa-miR-523, hsa-miR-524, hsa-miR-525, hsa-miR-526a-1, hsa-miR-526a-2, LEFTB, LHX1, LHX6, LIN28, MYBL2, MYC, MYON, NANOG, NFIX, OCT3/4 (POU5F1), OCT6 (POU3F1), OTX2, PHC1, SALL4, SOX2, TERF1, TERT, TGIF, VENTX2, ZIC2, ZIC3, ZICS, and ZNF206. 
     
     
         53 . The composition of  claim 52 , wherein the composition comprises six reprogramming factors. 
     
     
         54 . The composition of  claim 52 , wherein the reprogramming factors are selected from the group consisting of SOX2, NANOG, cMYC, OCT4, and DNMT3B. 
     
     
         55 . The composition of  claim 52 , wherein the reprogramming factors are proteins. 
     
     
         56 . The composition of  claim 52 , wherein the reprogramming factors are microRNAs. 
     
     
         57 . The composition of  claim 52 , wherein the composition comprises a cell-free extract obtained from undifferentiated cells. 
     
     
         58 . The composition of  claim 57 , wherein the cell-free extract is obtained from undifferentiated cells engineered to overexpress one or more reprogramming factors. 
     
     
         59 . The composition of  claim 57 , wherein the cell-free extract is supplemented with one or more reprogramming factors. 
     
     
         60 . The composition of  claim 57 , wherein the undifferentiated cells are embryonic stem cells. 
     
     
         61 . The composition of  claim 57 , further comprising a mammalian somatic cell. 
     
     
         62 . The composition of  claim 57 , further comprising a mammalian somatic cell nucleus. 
     
     
         63 . The composition of  claim 52 , wherein the composition comprises an enucleated cytoplast. 
     
     
         64 . The composition of  claim 63 , wherein the enucleated cytoplast is obtained from an undifferentiated cell engineered to overexpress one or more reprogramming factors. 
     
     
         65 . The composition of  claim 63 , wherein the enucleated cytoplast is supplemented with one or more reprogramming factors. 
     
     
         66 . The composition of  claim 63 , wherein the enucleated cytoplast is a cytoplast prepared from an embryonal carcinoma cell line. 
     
     
         67 . The composition of  claim 63 , wherein the enucleated cytoplast is a cytoplast prepared from an oocyte.