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

• US 20050283844A1
• Filed: 03/21/2005
• Published: 12/22/2005
• Est. Priority Date: 02/14/2001
• Status: Abandoned Application

• Current Assignee: Catherine M. Verfaillie, Leo T. Furcht, Morayma Reyes
• Original Assignee: Catherine M. Verfaillie, Leo T. Furcht, Morayma Reyes
• Inventors: Verfaillie, Catherine M., Furcht, Leo T., Reyes, Morayma
• Application Number:
  US11/084,809
• Publication Number:
  US 20050283844A1
• Time in Patent Office: Days
• US Class Current: 800/14
• CPC Class Codes: A01K 2217/05 Animals comprising random inserted nucleic acids (transgenic)
• CPC Class Codes: A01K 2217/075 inducing loss of function, i.e. knock out
• CPC Class Codes: A01K 2227/105 Murine
• CPC Class Codes: A01K 2227/106 Primate
• CPC Class Codes: A01K 67/0271 Chimeric vertebrates, e.g. comprising exogenous cells
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 39/001 Preparations to induce tolerance to non-self, e.g. prior to transplantation
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• CPC Class Codes: A61P 1/00 Drugs for disorders of the alimentary tract or the digestive system
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 13/10 of the bladder
• CPC Class Codes: A61P 13/12 of the kidneys
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 17/00 Drugs for dermatological disorders
• CPC Class Codes: A61P 19/04 for non-specific disorders of the connective tissue
• CPC Class Codes: A61P 19/08 for bone diseases, e.g. rachitism, Paget's disease
• CPC Class Codes: A61P 21/00 Drugs for disorders of the muscular or neuromuscular system
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/02 for peripheral neuropathies
• CPC Class Codes: A61P 27/02 : Ophthalmic agents
• CPC Class Codes: A61P 3/00 : Drugs for disorders of the metabolism of the blood or the extracellular fluid A61P7/00
• CPC Class Codes: A61P 3/10 : for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 31/00 : Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
• CPC Class Codes: A61P 31/04 : Antibacterial agents
• CPC Class Codes: A61P 31/10 : Antimycotics
• CPC Class Codes: A61P 31/12 : Antivirals
• CPC Class Codes: A61P 35/00 : Antineoplastic agents
• CPC Class Codes: A61P 37/00 : Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 : Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 39/02 : Antidotes
• CPC Class Codes: A61P 41/00 : Drugs used in surgical methods, e.g. surgery adjuvants for preventing adhesion or for vitreum substitution
• CPC Class Codes: A61P 43/00 : Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 : Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: A61P 7/04 : Antihaemorrhagics; Procoagulants; Haemostatic agents; Antifibrinolytic agents
• CPC Class Codes: A61P 7/06 : Antianaemics
• CPC Class Codes: A61P 7/08 : Plasma substitutes; Perfusion solutions; Dialytics or haemodialytics; Drugs for electrolytic or acid-base disorders, e.g. hypovolemic shock artificial tears A61P27/04
• CPC Class Codes: A61P 9/00 : Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/10 : for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 15/873 : Techniques for producing new embryos, e.g. nuclear transfer, manipulation of totipotent cells or production of chimeric embryos
• CPC Class Codes: C12N 2501/11 : Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/113 : Acidic fibroblast growth factor (aFGF, FGF-1)
• CPC Class Codes: C12N 2501/115 : Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/117 : Keratinocyte growth factors (KGF-1, i.e. FGF-7; KGF-2, i.e. FGF-12)
• CPC Class Codes: C12N 2501/119 : Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
• CPC Class Codes: C12N 2501/12 : Hepatocyte growth factor [HGF]
• CPC Class Codes: C12N 2501/135 : Platelet-derived growth factor [PDGF]
• CPC Class Codes: C12N 2501/235 : Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2501/237 : Oncostatin M [OSM]
• CPC Class Codes: C12N 2502/08 : cells of the nervous system
• CPC Class Codes: C12N 2502/30 : tumour cells
• CPC Class Codes: C12N 2503/00 : Use of cells in diagnostics
• CPC Class Codes: C12N 2506/03 : from non-embryonic pluripotent stem cells
• CPC Class Codes: C12N 2517/02 : Cells from transgenic animals
• CPC Class Codes: C12N 5/0607 : Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0619 : Neurons
• CPC Class Codes: C12N 5/0622 : Glial cells, e.g. astrocytes, oligodendrocytes; Schwann cells
• CPC Class Codes: C12N 5/067 : Hepatocytes

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.

Claims

1-101. -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.