Regulating stem cells

• US 10,358,629 B2
• Filed: 11/16/2015
• Issued: 07/23/2019
• Est. Priority Date: 03/08/2006
• Status: Active Grant

• Current Assignee: Kwalata Trading Ltd.
• Original Assignee: Kwalata Trading Ltd.
• Inventors: Porat, Yael, Fulga, Valentin, Porozov, Svetlana, Belleli, Adina
• Primary Examiner(s): Schuberg, Laura
• Application Number:
  US14/942,259
• Publication Number:
  US 20160145579A1
• Time in Patent Office: 1,345 Days
• Field of Search: None
• 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

Provided are methods for producing progenitor/precursor cells from a population of initiating cells (ICP) that have a density of less than 1.072 g/ml and at least 25% of which are CD31Bright by in vitro stimulating the ICP with different factors.

Claims

1. A method of making a composition, the method comprising:

obtaining 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 CD31^Bright; and

in vitro stimulating the ICP by culturing the ICP in the presence of one or more factors selected from the group consisting of;

autologous serum, bFGF, and IFN beta to differentiate into a progenitor/precursor cell population (PCP) comprising a subpopulation of cells that express one or more markers selected from the group consisting of;

connexin 43, alfa actin, IgG1, IgG2a, and troponin.

2. The method according to claim 1, comprising preparing the PCP as a product for administration to a patient.

3. The method according to claim 1, comprising freezing the ICP prior to stimulating the ICP.

4. The method according to claim 1, comprising freezing the PCP.

5. The method according to claim 1, comprising transporting the PCP to a site at least 10 km from a site where the PCP is first created.

6. The method according to claim 1, wherein stimulating the ICP comprises:

culturing the ICP in a first container during a first portion of a culturing period;

removing cells of the ICP from the first container at the end of the first portion of the period; and

culturing, in a second container during a second portion of the period, the cells removed from the first container.

7. The method according to claim 1, comprising locally administering the PCP to a site of the patient including injured tissue.

8. The method according to claim 7, wherein locally administering the PCP comprises implanting at the site a device including the PCP.

9. The method according to claim 8, comprising using the device to enable increased survival of PCP in injured tissue.

10. The method according to claim 8, comprising configuring the device for slow release of cells of the PCP into the injured tissue.

11. The method according to claim 8, comprising secreting, from the PCP, therapeutic molecules to the tissue.

12. The method according to claim 8, comprising secreting, from the device, soluble molecules that support the PCP.

13. A method of making a composition, the method comprising obtaining 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 CD31^Bright; and

in vitro stimulating the ICP by culturing the ICP in the presence of one or more factors selected from the group consisting of;

autologous serum, bFGF, IFN beta, erythropoietin, a statin, an antidiabetic agent, a thiazolidinedione, rosiglitazone, a proliferation-differentiation-enhancing agent, anti-CD34, anti-Tie-2, anti-CD133, anti-CD117, LIF, EPO, IGF, M-CSF, GM-CSF, TGF alpha, TGF beta, VEGF, BHA, BDNF, GDNF, NGF, NT3, NT4/5, S-100, CNTF, EGF, NGF3, CFN, ADMIF, estrogen, prolactin, an adrenocorticoid hormone, ACTH, MCT-165, glatiramer acetate, a glatiramer acetate-like molecule, IFN alpha, glutamate, serotonin, acetylcholine, NO, retinoic acid (RA), insulin, forskolin, and cortisone to differentiate into a progenitor/precursor cell population (PCP) comprising a subpopulation of cells that express one or more markers selected from the group consisting of connexin 43, alfa actin, IgG1, IgG2a and troponin.

14. A method of making a composition, the method comprising obtaining 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 CD31^Bright; and

in vitro stimulating the ICP by culturing the ICP in the presence of one or more factors selected from the group consisting of;

autologous serum, bFGF, IFN beta, anti-Tie-2, anti-CD133, and anti-CD117, VEGF, anti-VEGF, anti-VEGF receptor, heparin, MCDB-201, sodium selenite, dexamethasone, and BSA.