MULTIPOTENT STEM CELLS AND USES THEREOF

• US 20100291042A1
• Filed: 05/05/2008
• Published: 11/18/2010
• Est. Priority Date: 05/03/2007
• Status: Abandoned Application

• Current Assignee: Brigham and Women's Hospital Incorporated
• Original Assignee: Brigham and Women's Hospital Incorporated
• Inventors: Southgate, Christopher, Crawford, Keith W.
• Application Number:
  US12/598,047
• Publication Number:
  US 20100291042A1
• Time in Patent Office: Days
• US Class Current: 424/93.7
• 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 35/14 Blood; Artificial blood perfluorocarbons A61K31/02; umbilical cord blood A61K35/51; haemoglobin A61K38/42
• CPC Class Codes: A61K 35/24 Mucus; Mucous glands; Bursa; Synovial fluid; Arthral fluid; Excreta; Spinal fluid saliva A61K35/38
• CPC Class Codes: A61P 17/02 for treating wounds, ulcers, burns, scars, keloids, or the like
• CPC Class Codes: A61P 37/02 Immunomodulators
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: C12N 2500/25 Insulin-transferrin; Insulin-transferrin-selenium
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/38 Vitamins
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/125 Stem cell factor [SCF], c-kit ligand [KL]
• CPC Class Codes: C12N 2501/135 Platelet-derived growth factor [PDGF]
• CPC Class Codes: C12N 2501/155 Bone morphogenic proteins [BMP]; Osteogenins; Osteogenic factor; Bone inducing factor
• CPC Class Codes: C12N 2501/22 Colony stimulating factors (G-CSF, GM-CSF)
• CPC Class Codes: C12N 2501/2303 Interleukin-3 (IL-3)
• CPC Class Codes: C12N 2501/2306 Interleukin-6 (IL-6)
• CPC Class Codes: C12N 2501/235 Leukemia inhibitory factor [LIF]
• CPC Class Codes: C12N 2501/26 Flt-3 ligand (CD135L, flk-2 ligand)
• CPC Class Codes: C12N 2501/39 Steroid hormones
• CPC Class Codes: C12N 2501/40 : Regulators of development
• CPC Class Codes: C12N 2501/41 : Hedgehog proteins; Cyclopamine (inhibitor)
• CPC Class Codes: C12N 2501/999 : Small molecules not provided for elsewhere
• CPC Class Codes: C12N 5/0639 : Dendritic cells, e.g. Langherhans cells in the epidermis
• CPC Class Codes: C12N 5/0647 : Haematopoietic stem cells; Uncommitted or multipotent progenitors
• CPC Class Codes: C12N 5/0653 : Adipocytes; Adipose tissue
• CPC Class Codes: C12N 5/0654 : Osteocytes, Osteoblasts, Odontocytes; Bones, Teeth
• CPC Class Codes: C12N 5/0655 : Chondrocytes; Cartilage
• CPC Class Codes: C12N 5/0657 : Cardiomyocytes; Heart cells
• CPC Class Codes: C12N 5/0658 : Skeletal muscle cells, e.g. myocytes, myotubes, myoblasts
• CPC Class Codes: C12N 5/0661 : Smooth muscle cells
• CPC Class Codes: C12N 5/0662 : Stem cells
• CPC Class Codes: C12N 5/0668 : Mesenchymal stem cells from other natural sources
• CPC Class Codes: C12N 5/067 : Hepatocytes
• CPC Class Codes: C12N 5/0676 : Pancreatic cells

Abstract

The invention provides a quiescent stem cell having the capacity to differentiate into ectoderm, mesoderm and endoderm, and which does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105 and CD90. The invention further provides a proliferative stem cell, which expresses genes including Oct-4, Nanog, Sox2, GDF3, P16INK4, BMI, Notch, HDAC4, TERT, Rex-1 and TWIST but does not express cell surface markers including MHC class I, MHC class II, CD44, CD45, CD13, CD34, CD49c, CD73, CD105 and CD90. The cells of the invention can be isolated from adult mammals, have embryonic cell characteristics, and can form embryoid bodies. Methods for obtaining the stem cells, as well as methods of treating diseases and differentiated the stem cells, are also provided.

Claims

1. An isolated stem cell which is capable of proliferating and differentiating into ectoderm, mesoderm, and endoderm, expresses at least one of Oct-4, Nanog, Sox-2, Rex-1, GDF-3, and Stella and does not express CD13, CD44, CD45, CD90, and CD105.

3. The cell of claim 1, wherein said cell further does not express at least one of MHC class I, MHC class II, CD34, CD49c, and CD73.

4. The cell of claim 1, wherein said cell is isolated from synovial fluid, blood or tissue.

5. The cell of claim 1, wherein said cell is substantially purified.

6. The cell of claim 1, wherein said cell is isolated from a mammal.

7. The cell of claim 6, wherein said cell is isolated from a human.

8. The cell of claim 1, wherein said cell is isolated from an adult mammal.

9. The cell of claim 1, comprising a heterologous nucleic acid sequence.

10. The cell of claim 9, wherein said heterologous nucleic acid sequence comprises a stem cell-specific promoter.

11. The cell of claim 10, wherein said promoter is the promoter of an embryonic transcription factor.

12. The cell of claim 11, wherein said promoter is selected from the group consisting of the Oct-4 promoter, Nanog promoter, Sox2 promoter, cMyc-2 promoter, KLF4 promoter, Rex-1 promoter, GDF-3 promoter, Stella promoter, FoxD3 promoter, Polycomb Repressor Complex 2 promoter, and CTCF promoter.

13. The cell of claim 10, wherein said promoter is operably linked to a detectable gene product.

14. The cell of claim 13, wherein said detectable gene product is GFP.

15. A population of cells wherein at least 10% of the cells are the stem cells of claim 1.

17. A composition comprising:

(a) a population of cells of claim 15; and

(b) a cryoprotectant, a dendritic cell or an antigen presenting cell or a committed or differentiated cell.

38. A method of forming a muscle cell, said method comprising culturing a stem cell of claim 1 under muscle cell differentiating conditions.

40. A method of forming a neural cell, said method comprising culturing a stem cell of claim 1 under neural cell-forming conditions.

42. A method of forming a hematopoietic cell, the method comprising culturing a stem cell of claim 1 under hematopoietic cell-forming conditions.

44. A method of forming an endothelial cell, the method comprising culturing a stem cell of claim 1 under endothelial cell-forming conditions.

46. A method of forming a hematopoietic cell, the method comprising culturing a stem cell of claim 1 under hematopoietic cell-forming conditions.

49. A method for promoting wound healing in a subject, said method comprising administering a stem cell of claim 1, or a committed or differentiated progeny thereof, to said wound or to a site near said wound in an amount sufficient to promote the healing of said wound.

51. A method for treating a cardiovascular disease in a subject, said method comprising administering to said subject a stem cell of claim 1, or a committed or differentiated progeny thereof, in an amount sufficient to treat said disease.

53. A method of increasing vascularization in a subject, said method comprising administering to said subject a stem cell of claim 1, or a committed or differentiated progeny thereof, in an amount sufficient to increase vascularization.

55. A method for treating a neurological disorder in a subject, said method comprising administering to said subject a stem cell of claim 1, or a committed or differentiated progeny thereof, in an amount sufficient to treat said disease.

58. A method for treating an autoimmune disease in a subject, said method comprising administering to said subject a stem cell of claim 1, or a committed or differentiated progeny thereof, in an amount sufficient to treat said disease.

59. A method for reducing or preventing rejection of transplanted tissue in a subject, said method comprising administering to said subject a stem cell of claim 1 or a committed or differentiated progency thereof, in an amount sufficient to reduce or prevent said rejection.

2. An isolated quiescent stem cell which is capable of proliferating and differentiating into ectoderm, mesoderm, and endoderm and does not express Oct-4, CD13, CD44, CD45, CD90, and CD105.

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21. A method of isolating a population of stem cells, said method comprising the steps:

(a) providing a bodily fluid or tissue from a subject;

(b) enriching for a population of cells that are about 6 μ

m to 20 μ

m in size; and

(c) depleting cells from said population expressing a stem cell surface marker or an MHC protein, thereby isolating a population of stem cells.

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