Regulating Stem Cells

• US 20100291610A1
• Filed: 03/08/2007
• Published: 11/18/2010
• Est. Priority Date: 03/08/2006
• Status: Active Grant

• Current Assignee: Kwalata Trading Ltd.
• Original Assignee: Kwalata Trading Ltd.
• Inventors: Belleli, Adina, Porozov, Svetlana, Porat, Yael, Fulga, Valentin
• Granted Patent: US 8,541,232 B2
• Time in Patent Office: Days
• US Class Current: 435/29
• CPC Class Codes: A61K 2035/124 the cells being hematopoietic, bone marrow derived or blood cells
• CPC Class Codes: A61K 35/44 Vessels; Vascular smooth muscle cells; Endothelial cells; Endothelial progenitor cells
• CPC Class Codes: C12N 2501/14 Erythropoietin [EPO]
• CPC Class Codes: C12N 2501/165 Vascular endothelial growth factor [VEGF]
• CPC Class Codes: C12N 2501/23 Interleukins [IL]
• CPC Class Codes: C12N 2501/33 Insulin together with transferrin C12N2500/25; Insulin-like growth factors C12N2501/105
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2501/392 Sexual steroids
• CPC Class Codes: C12N 2501/599 with CD designations not provided for elsewhere
• CPC Class Codes: C12N 2501/91 Heparin
• CPC Class Codes: C12N 2506/11 from blood or immune system cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 2533/52 Fibronectin; Laminin
• CPC Class Codes: C12N 2533/54 Collagen; Gelatin
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0634 Cells from the blood or the immune system
• CPC Class Codes: C12N 5/0692 Stem cells; Progenitor cells; Precursor cells

Abstract

A composition of matter is provided, comprising a population of cultured cells that comprises a sub-population of cells that both stain as CD31Bright and demonstrate uptake of Ac-LDL+, and secrete IL-8. A method is also provided, comprising stimulating in vitro an initiating cell population (ICP) of at least 5 million cells that have a density of less than 1.072 g/ml, wherein at least 1% of the cells of the ICP is CD34+CD45−/Dim, and at least 25% of the cells of the ICP are CD31Bright, to differentiate into a progenitor/precursor cell population (PCP). Other embodiments are also described.

Claims

1-14. -14. (canceled)

15. A composition of matter, comprising a population of cultured cells that comprises a sub-population of cells that stain as CD31Bright, demonstrate uptake of Ac-LDL+ and secrete interleukin-8.

16. The composition of matter according to claim 15, wherein the sub-population comprises at least 10% of the cells in the population.

19. The composition of matter according to claim 15, wherein the sub-population secretes at least 50 pg IL-8 per 10⁶ cells/ml over a period of at least 24 hours.

160. The composition of matter according to claim 15, wherein at least 1.5% of the cells of the population comprise a feature selected from the group consisting of:

CD34, CD117, CD133, Tie-2, CD34+CD133+, KDR, CD34+KDR+, CD144, von Willebrand Factor, SH2 (CD105), SH3, fibronectin, collagen type I, collagen type III, collagen type IV, ICAM type 1, ICAM type 2, VCAM1, vimentin, BMP-R IA, BMP-RII, CD44, integrin b1, aSM-actin, MUC18, CXCR4 and CXCR8.

161. The composition of matter according to claim 15, wherein at least 1.5% of the cells of the population secrete a molecule selected from the group consisting of:

angiogenin, VEGF, MMP2, and MMP9.

162. The composition of matter according to claim 15, wherein at least 1.5% of the cells of the population include a feature selected from the group consisting of:

a tube-like structure, a tendency to form a colony, a tendency to form a cluster, and a tendency to migrate toward a chemoattractant selected from the group consisting of;

bFGF, VEGF, SCF, G-CSF, GM-CSF, SDF-1, and IL-8.

17-18. -18. (canceled)

20-159. -159. (canceled)

163. A method comprising:

providing an initiating cell population (ICP) of at least 5 million cells that have a density of less than 1.072 g/ml, at least 1% of which are CD34+CD45−

/Dim, and at least 25% of which are CD31Bright; and

stimulating the ICP to differentiate into a progenitor/precursor cell population (PCP).

164. The method according to claim 163, wherein the ICP includes at least 5 million cells, and wherein stimulating the ICP comprises stimulating the ICP that includes the at least 5 million cells.

165. The method according to claim 163, wherein at least 1.5% of the cells of the ICP are CD34+CD45−

/Dim, and wherein stimulating the ICP comprises stimulating the ICP of which at least 1.5% of the cells are CD34+CD45−

/Dim.

166. The method according to claim 163, wherein at least 30% of the cells of the ICP are CD31Bright, and wherein stimulating the ICP comprises stimulating the ICP of which at least 30% of the cells are CD31Bright.

167. The method according to claim 163, wherein the ICP includes at least 5 million cells that have a density of less than 1.062 g/ml, at least 1% of which are CD34+CD45−

/Dim, and wherein stimulating the ICP comprises stimulating the ICP that has the at least 5 million cells that have a density of less than 1.062 g/ml.

168. The method according to claim 163, further comprising preparing the PCP as a product for administration to a patient.

169. The method according to claim 163, wherein the ICP is characterized by at least 2.5% of the ICP being CD34+CD45−

/Dim, and wherein stimulating the ICP comprises stimulating the ICP having the at least 2.5% of the ICP that are CD34+CD45−

/Dim.

170. The method according to claim 163, wherein stimulating the ICP comprises culturing the ICP for a period lasting between 1 and 5 days in a culture medium comprising less than or equal to 10% serum.

171. The method according claim 163, wherein stimulating the ICP comprises culturing the ICP in the presence of IL-8.

172. The method according to claim 163, further comprising identifying the PCP as being suitable for therapeutic implantation in response to an assessment that at least 1.5% of cells of the PCP demonstrate a feature selected from the group consisting of:

a desired morphology, a desired cellular marker, a desired cellular component, a desired enzyme, a desired receptor, a desired genotypic feature, and a desired physiological feature.

173. The method according claim 163, further comprising identifying the PCP as being suitable for therapeutic implantation in response to an assessment that the PCP includes at least 1 million angiogenic cell precursors (ACPs).

174. The method according to claim 163, further comprising identifying the PCP as being suitable for therapeutic implantation in response to an assessment that the PCP includes at least 1 million cardiomyocyte progenitors.

175. The method according to claim 163, further comprising identifying the PCP as being suitable for therapeutic implantation in response to an assessment that the PCP includes at least 1 million neural cell progenitors.

176. The method according claim 163, further comprising characterizing the PCP as including angiogenic cell precursors (ACPs), in response to an evaluation of at least one feature selected from the group consisting of:

a phenotypical feature of cells in the PCP, a genotypical feature of cells in the PCP, and a physiological feature of cells in the PCP.