Multipotent adult stem cells, sources thereof, methods of obtaining and maintaining same, methods of differentiation thereof, methods of use thereof and cells derived thereof

Abstract

Methods and compositions are provided for circularizing target sequences in a sample. In particular, ligation oligonucleotides are employed to selectively hybridize with the target such that the target can be ligated into a closed circular target. Rolling circle amplification can then be performed directly on the target sequence for subsequent detection and analysis.

1 - 101 . (canceled)  
     
     
         102 . A substantially homogenous cell population which co-expresses CD49c, CD90 and at least one cardiac-related transcription factor.  
     
     
         103 . The substantially homogenous cell population of  claim 102 , further including co-expression of telomerase.  
     
     
         104 . The substantially homogenous cell population of  claim 102 , wherein the cells are derived from human bone marrow cells.  
     
     
         105 . The substantially homogenous cell population of  claim 102 , wherein the cardiac-related transcription factor is selected from the group consisting of GATA4, Irx4 and Nkx2.5.  
     
     
         106 . The substantially homogenous cell population of  claim 102 , further including a label.  
     
     
         107 . The substantially homogenous cell population of  claim 102 , wherein the cell population differentiates into cardiac muscle cells.  
     
     
         108 . The substantially homogenous cell population of  claim 102 , wherein the cells express at least one trophic factor selected from the group consisting of IL-6, VEGF, MCP1 and BDNF.  
     
     
         109 . A substantially homogenous cell population which co-expresses CD49c, CD90, and at least one cardiac-related transcription factor, but does not express bone sialoprotein.  
     
     
         110 . The substantially homogenous cell population of  claim 109 , wherein the cardiac-related transcription factor is selected from the group consisting of GATA4, Irx4 and Nkx2.5.  
     
     
         111 . A substantially homogenous cell population which co-expresses CD49c, CD90, GATA4, Irx4 and Nkx2.5.  
     
     
         112 . A substantially homogenous cell population which co-expresses CD49c, CD90, telomerase, GATA4, Irx4 and Nkx2.5.  
     
     
         113 . A method of making a substantially homogenous cell population which co-expresses CD49c, CD90 and at least one cardiac-related transcription factor, comprising the steps of: a) culturing a source of the cell population under a low oxygen condition; and b) treating the cultured source of the cell population with a protein kinase C inhibitor and a DNA methylation inhibitor.  
     
     
         114 . The method of  claim 113 , wherein the source of the cell population includes a bone marrow source.  
     
     
         115 . The method of  claim 113 , wherein the protein kinase C inhibitor is chelerythrine.  
     
     
         116 . The method of  claim 115 , wherein the DNA methylation inhibitor is 5-azacytidine.  
     
     
         117 . The method of  claim 113 , wherein the treated cell population co-expresses a cardiac-related transcription factor selected from the group consisting of GATA4, Irx4 and Nkx2.5.  
     
     
         118 . A method of making a substantially homogenous cell population which co-expresses CD49c, CD90, telomerase and at least one cardiac-related transcription factor, comprising the steps of: a) culturing a source of the cell population under a low oxygen condition; and b) treating the cultured source of the cell population with a protein kinase C inhibitor and a DNA methylation inhibitor.  
     
     
         119 . A method of making a substantially homogenous cell population which co-expresses CD49c, CD90, telomerase, GATA4, Irx4 and Nkx2.5, comprising the steps of: a) culturing a source of the cell population under a low oxygen condition; and b) treating the cultured source of the cell population with a protein kinase C inhibitor and a DNA methylation inhibitor.  
     
     
         120 . The method of  claim 119 , wherein the source of the cell population includes a bone marrow source.  
     
     
         121 . The method of  claim 119 , wherein the protein kinase C inhibitor is chelerythrine.  
     
     
         122 . The method of  claim 121 , wherein the DNA methylation inhibitor is 5azacytidine.  
     
     
         123 . A method of making a substantially homogenous cell population which co-expresses CD49c, CD90, GATA4, Irx4 and Nkx2.5, comprising the steps of: a) culturing a source of the cell population under a low oxygen condition; and b) treating the cultured source of the cell population with a protein kinase A inhibitor and a DNA methylation inhibitor.  
     
     
         124 . A method of making a substantially homogenous cell population which co-expresses CD49c, CD90 and at least one cardiac-related transcription factor, comprising the step of treating a cell population which co-expresses CD49c and CD90 with a protein kinase C inhibitor and a DNA methylation inhibitor.  
     
     
         125 . A method of making a substantially homogenous cell population which co-expresses CD49c, CD90, telomerase and at least one cardiac-related transcription factor, comprising the step of treating a cell population which co-expresses CD49c, CD90 and telomerase with a protein kinase C inhibitor and a DNA methylation inhibitor.  
     
     
         126 . A method of making a substantially homogenous cell population which co-expresses CD49c, CD90, telomerase, GATA4, Irx4 and Nkx2.5, comprising the step of treating a cell population which co-expresses CD49c, CD90 and telomerase with a protein kinase C inhibitor and a DNA methylation inhibitor.  
     
     
         127 . A method of treating a myocardial infarction in a human, comprising the step of administering a substantially homogenous cell population which co-expresses CD49c, CD90 and at least one cardiac-related transcription factor to the human.  
     
     
         128 . The method of  claim 127 , wherein the cardiac-related transcription factor is selected from the group consisting of GATA4, Irx4 and Nkx2.5.  
     
     
         129 . A method of treating a myocardial infarction in a human comprising the step of administering a substantially homogenous cell population which co-expresses CD49c, CD90 and at least one cardiac-related transcription factor to the human.  
     
     
         130 . A method of treating a myocardial infarction in a human comprising the step of administering to the human a substantially homogenous cell population which co-expresses CD49c, CD90, GATA4, Irx4 and Nkx2.5.  
     
     
         131 . A method of treating a myocardial infarction in a human, comprising the step of administering to the human a substantially homogenous cell population which co-expresses CD49c, CD90, telomerase, GATA4, Irx4 and Nkx2.5.  
     
     
         132 . A method of treating a myocardial infarction in a human, comprising the steps of: a) culturing a source of a cell population under a low oxygen condition; b) treating the cultured source of the cell population with a protein kinase C inhibitor and a DNA methylation inhibitor; and d) administering the treated cell population to the human.  
     
     
         133 . The method of  claim 132 , wherein the treated cell population is administered proximate to the myocardial infarction.  
     
     
         134 . The method of  claim 133 , wherein the treated cell population is administered into a cardiac muscle.  
     
     
         135 . The method of  claim 132 , further including selecting from the treated cell population, a population of cells which co-expresses CD49c, CD90, and at least one cardiac-specific marker.  
     
     
         136 . The method of  claim 135 , wherein the selected cell population further includes cells which express telomerase.  
     
     
         137 . The method of  claim 135 , wherein the cardiac-specific marker is selected from the group consisting of GATA4, Irx4 and Nkx2.5.  
     
     
         138 . The method of  claim 132 , wherein the source of the cell population includes a bone marrow source.  
     
     
         139 . A method of treating a myocardial infarction in a human, comprising the steps of: a) treating a cell population which co-expresses CD49c and CD90 with a protein kinase C inhibitor and a DNA methylation inhibitor; and b) administering the treated cells to the human.  
     
     
         140 . The method of  claim 139 , wherein the cell population is derived from bone marrow.  
     
     
         141 . The method of  claim 139 , wherein the cell population further includes cells which co-express telomerase.  
     
     
         142 . The method of  claim 139 , wherein the cell population expresses at least one cardiac-related transcription factor selected from the group consisting of GATA4, Irx4 and Nkx2.5.  
     
     
         143 . A method of treating a congestive heart failure in a human, comprising the step of administering a substantially homogenous cell population which co-expresses CD49c, CD90 and at least one cardiac-related transcription factor to the human.  
     
     
         144 . A method of treating a congestive heart failure in a human comprising the step of administering a substantially homogenous cell population which co-expresses CD49c, CD90 and at least one cardiac-related transcription factor to the human.  
     
     
         145 . A method of treating a congestive heart failure in a human comprising the step of administering a substantially homogenous cell population which co-expresses CD49c, CD90, GATA4, Irx4 and Nkx2.5 to the human.  
     
     
         146 . A method of treating a congestive heart failure in a human, comprising the step of administering to the human a substantially homogenous cell population which co-expresses CD49c, CD90, telomerase, GATA4, Irx4 and Nkx2.5.  
     
     
         147 . A method of treating a congestive heart failure in a human, comprising the steps of: a) culturing a source of a cell population under a low oxygen condition; b) treating the cultured source of the cell population with a protein kinase C inhibitor and a DNA methylation inhibitor; and d) administering the treated cell population to the human  
     
     
         148 . A method of forming a committed progenitor cell-type, comprising the step of combining a substantially homogenous population of cells that co-expresses CD49c and CD90 with a population of cells that includes at least one committed progenitor cell type.  
     
     
         149 . The method of  claim 148 , wherein said population of cells is selected from the group consisting of a population of nerve cells and a population of cardiac muscle cells.  
     
     
         150 . The method of  claim 148 , wherein the population of cells expresses telomerase.  
     
     
         151 . A substantially homogenous cell population which co-expresses CD49c, CD90 and has a doubling time of less that about 144 hours when cultured under a low oxygen condition.  
     
     
         152 . The substantially homogenous cell population of  claim 151 , wherein the doubling time is less than about 72 hours.  
     
     
         153 . The substantially homogenous cell population of  claim 151 , wherein the doubling time is less than about 48 hours.  
     
     
         154 . The substantially homogenous cell population of  claim 151 , wherein the doubling time is less than about 65 hours.  
     
     
         155 . The substantially homogenous cell population of  claim 151 , wherein the doubling time is less than about 35 hours.  
     
     
         156 . The substantially homogenous cell population of  claim 151 , wherein the low oxygen condition is less than about 5% oxygen.  
     
     
         157 . A substantially homogenous cell population which co-expresses CD49c, CD90 and has a doubling time less than about 144 hours when cultured under a low oxygen condition, wherein the substantially homogenous cell population is formed by a method, comprising the step of culturing a cell population source at a seeding density of about 100 cells/cm.sup.2 under the low oxygen condition.  
     
     
         158 . A pharmaceutical composition comprising a substantially homogenous cell population which co-expresses CD49c, CD90 and at least one cardiac-related transcription factor.  
     
     
         159 . A pharmaceutical composition comprising a substantially homogenous cell population which co-expresses CD49c, CD90, telomerase and at least one cardiac-related transcription factor.  
     
     
         160 . A pharmaceutical composition comprising a substantially homogenous cell population which co-expresses CD49c, CD90, telomerase, GATA4, Irx4 and Nkx2.5.  
     
     
         161 . A pharmaceutical composition comprising a substantially homogenous cell population which co-expresses CD49c, CD90, GATA4, Irx4 and Nkx2.5.