Multipotent stem cells and uses thereof

• US 9,127,252 B2
• Filed: 03/13/2013
• Issued: 09/08/2015
• Est. Priority Date: 05/03/2007
• Status: Active Grant

• Current Assignee: Brigham and Women's Hospital Incorporated
• Original Assignee: Brigham and Women's Hospital Incorporated
• Inventors: Crawford, Keith D., Southgate, Christopher
• Primary Examiner(s): DRISCOLL, LORA E BARNHART
• Application Number:
  US13/798,720
• Publication Number:
  US 20130336934A1
• Time in Patent Office: 909 Days
• Field of Search: None
• US Class Current: 1/1
• 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 the differentiated stem cells, are also provided.

Claims

1. A method for promoting wound healing in a subject, said method comprising administering to a human subject an effective dose of a formulation comprising:

(i) a population of stem cells that express a stem cell transcription factor but do not detectably express MHC Class I or cell surface markers CD44, CD45, and CD90, and

(ii) a pharmaceutically acceptable carrier,

in an amount sufficient to promote or improve the healing of said wound.

10. The method of claim 1, wherein the stem cells do not detectably express CD13 or CD105.

2. A method for promoting wound healing in a subject, said method comprising administering a formulation comprising:

(i) a population of stem cells that express a stem cell transcription factor but do not detectably express MHC Class I or cell surface CD44, CD45, and CD90,

(ii) at least one subpopulation of differentiated progeny cells, and

(iii) a pharmaceutically acceptable carrier,

in an amount sufficient to promote or improve the healing of said wound.

11. The method of claim 2, wherein the stem cells do not detectably express CD13 or CD105.

3. A method for promoting tissue generation in a subject, said method comprising transplanting into a subject at a surgical site a formulation comprising:

(i) a population of stem cells that express a stem cell transcription factor but do not detectably express MHC Class I or cell surface markers CD44, CD45, and CD90, and

(ii) a pharmaceutically acceptable carrier,

wherein the population of stem cells engrafts to target tissue, reduces inflammation in the target tissue and differentiates in vivo to match the tissue type and supplement the target tissue, thereby promoting tissue generation in the subject.

4. The method of claim 3, where the surgical site is the site of a joint procedure.

5. The method of claim 4, where the surgical site is the site of repair of articular cartilage.

6. The method of claim 5, wherein transplanting is by implanting or injecting the formulation into a patient'"'"'s injured cartilage tissue.

7. The method of claim 3, where the surgical site is the site of a cosmetic procedure.

12. The method of claim 3, wherein the stem cells do not detectably express CD13 or CD105.

8. A method for promoting tissue generation in a subject, said method comprising transplanting into a subject at a surgical site a formulation comprising:

(i) a population of stem cells that express a stem cell transcription factor but do not detectably express MHC Class I or cell surface markers CD44, CD45, and CD90,

(ii) a cellular differentiating agent, and

(iii) a pharmaceutically acceptable carrier,

wherein the population of stem cells engrafts to target tissue, reduces inflammation in the target tissue and differentiates in vivo to match the tissue type and supplement the target tissue, thereby promoting tissue generation in the subject.

9. The method of claim 8, where the surgical site is the site of an orthopedic, bone or cosmetic procedure.

13. The method of claim 8, wherein the stem cells do not detectably express CD13 or CD105.