Multipotent adult stem cells

• US 20090130065A1
• Filed: 04/12/2006
• Published: 05/21/2009
• Est. Priority Date: 04/12/2005
• Status: Active Grant

• Current Assignee: The Wistar Institute
• Original Assignee: Trustees Of The University Of Pennsylvania (University of Pennsylvania)
• Inventors: Yu, Hong, Xu, Xiaowei, Fang, Douglas Dong, Herlyn, Meenhard
• Granted Patent: US 8,999,706 B2
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: C12N 2500/44 Thiols, e.g. mercaptoethanol
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 5/0607 Non-embryonic pluripotent stem cells, e.g. MASC induced pluripotent stem cells C12N5/0696
• CPC Class Codes: C12N 5/0628 Hair stem cells; Hair progenitors mesenchymal stem cells from hair follicles C12N5/0666

Abstract

Isolated human multipotent adult stem cell and isolated populations of cells that include human multipotent adult stem cells are disclosed. Human hair-follicle derived multipotent adult stem cells and methods of preparing isolated populations of cells that include human multipotent adult stem cells are disclosed. Isolated human hair-follicle derived multipotent adult stem cell that can differentiate in culture into a neuronal cell, a glial cell, a melanocyte cell, a muscle cell, an osteocyte, a chondrocyte, and a lymphocyte. Isolated human hair-follicle derived multipotent adult stem cell that can grow in cell culture in spheres are disclosed. Human pancreas derived multipotent adult stem cells, human liver derived multipotent adult stem cells, human kidney derived multipotent adult stem cells, human heart derived multipotent adult stem cells, human neural derived multipotent adult stem cells and methods of preparing isolated populations of cells that include such human multipotent adult stem cells are disclosed. Method of treating an individual who has diabetes, cardiac muscle damage, muscle damage and disease, neurodegenerative disease or nerve damage or injury, bone loss, damage and/or disease, cartilage loss, damage and/or disease, hair loss and immune disorders, are disclosed.

Claims

1. An isolated human multipotent adult stem cell selected from the group consisting of:

an isolated human hair-follicle derived multipotent adult stem cell having one or more characteristics selected from the group consisting of;

expresses one or more of;

nanog, Oct4 and Nestin;

can differentiate in culture into one or more of the following;

a neuronal cell, a glial cell, a melanocytic cell, a muscle cell, an adipocyte, an osteocyte, a chondrocyte, an islet cell, and a lymphocyte; and

grows in cell culture in spheres; and

an isolated human multipotent adult stem cell selected from the group consisting of;

an isolated human multipotent adult stem cell that expresses nanog;

an isolated human multipotent adult stem cell that expresses Oct4 and is free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4; and

an isolated human multipotent adult stem cell that expresses nanog and Oct4 and is free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4.

2. The isolated human hair-follicle derived multipotent adult stem cell of claim 1 wherein the cell expresses a) one or more of:

nanog, Oct4 and Nestin, and b) is free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4.

3. The isolated human hair-follicle derived multipotent adult stem cell of claim 1 that expresses one or more of:

nanog, Oct4 and Nestin, wherein said cell expresses nanog and Oct4.

4. The isolated human hair-follicle derived multipotent adult stem cell of claim 3 wherein the cell is free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4.

5-7. -7. (canceled)

8. An isolated population of human multipotent adult stem cells selected from the group consisting of:

a) human multipotent adult stem cells that express nanog and is free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4;

human multipotent adult stem cells that express Oct4 and is free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4;

human multipotent adult stem cells that express Nestin and is free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4;

human multipotent adult stem cells that express nanog and Oct4 and is free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4; and

combinations thereof;

b) human hair-follicle derived multipotent adult stem cells that express nanog;

human hair follicle-derived multipotent adult stem cells that express Oct4;

human hair follicle-derived multipotent adult stem cells that express Nestin;

human hair-follicle derived multipotent adult stem cells that express nanog and Oct4; and

combinations thereof;

c) human hair-follicle derived multipotent adult stem cells that can differentiate in culture into a neuronal cell, a glial cell, a melanocytic cell, a muscle cell, an adipocyte, an osteocyte, a chondrocyte, an islet cell, and a lymphocyte;

d) human hair-follicle derived multipotent adult stem cell that grows in cell culture in spheres; and

e) isolated cells formed by the method of forming a single cell suspension of cells disassociated from a sample that comprises cells from an individual, and culturing such cells in conditions suitable for culturing and maintaining human embryonic stem cells or in conditions comprising a serum free minimal medium supplemented with human basic fibroblast growth factor and human epidermal growth factor;

wherein multipotent adult stem cells are identified and isolated from said culture.

10. The isolated population of human multipotent adult stem cells of claim 8 selected from the group consisting of:

human hair-follicle derived multipotent adult stem cells that express nanog;

human hair follicle-derived multipotent adult stem cells that express Oct4;

human hair follicle-derived multipotent adult stem cells that express Nestin;

human hair-follicle derived multipotent adult stem cells that express nanog and Oct4; and

combinations thereof,

wherein cells are free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4.

42. The isolated population of human multipotent adult stem cells of claim 8 wherein the cells are isolated cells formed by the method of forming a single cell suspension of cells disassociated from a sample that comprises cells from an individual, and culturing such cells in conditions suitable for culturing and maintaining human embryonic stem cells or in conditions comprising a serum free minimal medium supplemented with human basic fibroblast growth factor and human epidermal growth factor;

wherein multipotent adult stem cells are identified and isolated from said culture and said sample is selected from the group consisting of;

cells from follicular epithelium of a hair shaft;

a sample of liver tissue;

a sample of kidney, tissue;

a sample of pancreatic tissue;

a sample of cardiac tissue; and

a sample comprising nerve cells.

43. The isolated population of human multipotent adult stem cells of claim 8 wherein the cells are isolated cells formed by the method of forming a single cell suspension of cells disassociated from a sample that comprises cells from an individual, and culturing such cells in conditions suitable for culturing and maintaining human embryonic stem cells or in conditions comprising a serum free minimal medium supplemented with human basic fibroblast growth factor and human epidermal growth factor;

wherein multipotent adult stem cells are identified and isolated from said culture and said sample is cells from follicular epithelium of a hair shaft

9. (canceled)

11-12. -12. (canceled)

13. A method of preparing an isolated population of human hair-follicle derived multipotent adult stem cells comprising the steps of:

forming a single cell suspension of cells disassociated from follicular epithelium of a hair shaft and culturing such cells in conditions suitable for culturing and maintaining human embryonic stem cells wherein spheres form that comprise human hair-follicle derived multipotent adult stem cells or in conditions comprising a serum free minimal medium supplemented with human basic fibroblast growth factor and human epidermal growth factor;

or

culturing a human hair follicle in conditions suitable for culturing and maintaining human embryonic stem cells wherein spheres form that comprise human hair-follicle derived multipotent adult stem cells or in conditions comprising a serum free minimal medium supplemented with human basic fibroblast growth factor and human epidermal growth factor, wherein said hair follicle is cultured under such conditions for a time sufficient for cells to grow on said hair follicle, and isolating from said cells a population of human hair-follicle derived multipotent adult stem cells.

14-22. -22. (canceled)

23. A method of treating an individual comprising the steps selected from the group consisting of:

a) administering to said individual human isolated human hair-follicle derived multipotent adult stem cells having one or more characteristics selected from the group consisting of;

expresses one or more of;

nanog, Oct4 and Nestin;

can differentiate in culture into one or more of the following;

a neuronal cell, a glial cell, a melanocytic cell, a muscle cell, an adipocyte, an osteocyte, a chondrocyte, an islet cell, and a lymphocyte; and

grows in cell culture in spheres;

b) administering to said individual an isolated human multipotent adult stem cells selected from the group consisting of;

human multipotent adult stem cells that express nanog;

human multipotent adult stem cells that express Oct4 and are free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4;

human hair-follicle derived multipotent adult stem cells that express nanog and Oct4 and are free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4; and

human hair-follicle derived multipotent adult stem cells that can differentiate in culture into a neuronal cell, a glial cell, a melanocytic cell, a muscle cell, an adipocyte, an osteocyte, a chondrocyte, an islet cell, and a lymphocyte; and

human hair-follicle derived multipotent adult stem cell that grows in cell culture in spheres; and

c) administering to said individual human isolated population of multipotent adult stem cells having cells selected from the group consisting of;

human multipotent adult stem cells that express nanog and are free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4;

human multipotent adult stem cells that express Oct4 and are free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4;

human multipotent adult stem cells that express Nestin and are free of stage specific embryonic antigen-3 and stage specific embryonic antigen-4;

human multipotent adult stem cells that express nanog and Oct4 and are free of stage specific embryonic antigen-3 and stage specific embryonic antigen-combinations thereof,

d) administering to said individual human isolated population of human hair-follicle derived multipotent adult stem cells having cells selected from the group consisting of;

human hair-follicle derived multipotent adult stem cells that express nanog;

human hair follicle-derived multipotent adult stem cells that express Oct4;

human hair follicle-derived multipotent adult stem cells that express Nestin;

human hair-follicle derived multipotent adult stem cells that express nanog and Oct4; and

combinations thereof;

e) administering to said individual human isolated population of human hair-follicle derived multipotent adult stem cells having cells can differentiate in culture into one or more of the following;

a neuronal cell, a glial cell, a melanocytic cell, a muscle cell, an adipocyte, an osteocyte, a chondrocyte, an islet cell, and a lymphocyte; and

f) administering to said individual human isolated population of human hair-follicle derived multipotent adult stem cells having cells that grow in cell culture in spheres;

g) administering to said individual human isolated population of human hair-follicle derived multipotent adult stem cells formed by the method of forming a single cell suspension of cells disassociated from a sample that comprises cells from an individual and culturing such cells in conditions suitable for culturing and maintaining human embryonic stem cells or in conditions comprising a serum free minimal medium supplemented with human basic fibroblast growth factor and human epidermal growth factor;

wherein multipotent adult stem cells are identified and isolated from said culture and wherein said sample consists of cells from the follicular epithelium of a hair shaft a sample of liver tissue a sample of pancreatic tissue a sample of cardiac tissue and a sample of nerve cells.

and

h) administering to said individual isolated cells isolated by preparing an isolated population of human multipotent adult stem cells by forming a single cell suspension of cells disassociated from sample that comprises cells from an individual, and culturing such cells in conditions suitable for culturing and maintaining human embryonic stem cells or in conditions comprising a serum free minimal medium supplemented with human basic fibroblast growth factor and human epidermal growth factor;

wherein multipotent adult stem cells are identified and isolated from said culture.

25. The method of claim 23 wherein the individual has a disease or condition selected from the group consisting of:

hair loss;

liver damage or failure, pancreatic damage or failure, cardiac damage or failure;

neurodegenerative diseases;

bone loss, damage and/or disease;

cartilage loss, damage and/or disease.

44. The method of claim 23 wherein the individual has a disease or condition selected from the group consisting of:

diabetes;

cardiac muscle damage;

muscle damage or disease;

neurodegenerative disease;

nerve damage or injury;

hair loss;

bone loss;

damage and/or disease;

cartilage loss, damage and/or disease;

wherein

if the individual has diabetes, cells are administered to said individual, wherein the stem cells differentiate into insulin producing cells;

or cells are induced to differentiate into insulin producing cells and said insulin producing cells are administered to said individual;

if the individual has cardiac muscle damage, cells are administered to said individual'"'"'s heart, wherein the stem cells differentiate into muscle cells or muscle precursor cells;

or cells are induced to differentiate into muscle cells or muscle precursor cells and said muscle cells or muscle precursor cells are administered to said individual'"'"'s heart;

if the individual has muscle damage or disease, cells are administered to said individual'"'"'s muscle, wherein the stem cells differentiate into muscle cells or muscle precursor cells;

or cells are induced to differentiate into muscle cells or muscle precursor cells and said muscle cells or muscle precursor cells are administered to said individual'"'"'s muscle;

if the individual has neurodegenerative disease, cells are administered to said individual, wherein the stem cells differentiate into nerve cells;

or cells are induced to differentiate into nerve cells and said nerve cells are administered to said individual;

if the individual has nerve damager or injury, cells are administered to said individual, wherein the stem cells differentiate into nerve cells;

or cells are induced to differentiate into nerve cells and said nerve cells are administered to said individual;

if the individual has hair loss, cells are administered to said individual, wherein the stem cells differentiate into hair-producing cells or hair-producing precursor cells;

or cells are induced to differentiate into hair-producing cells or hair-producing precursor cells and hair-producing cells or hair-producing precursor cells are administered to said individual;

if the individual has bone loss, damage and/or disease, cells are administered to said individual, wherein the stem cells differentiate into osteocytes or osteocytes precursor cells;

or

cells are induced to differentiate into osteocytes or osteocytes precursor cells and said osteocytes or osteocytes precursor cells are administered to said individual;

if the individual has cartilage loss, damage and/or disease, cells are administered to said individual, wherein the stem cells differentiate into chondrocytes or chondrocytes precursor cells;

or cells are induced to differentiate into chondrocytes or chondrocytes precursor cells and said chondrocytes or chondrocytes precursor cells are administered to said individual;

45. The method of treating an individual of claim 23 comprising the step of:

administering to said individual, isolated cells isolated by preparing an isolated population of human multipotent adult stem cells by forming a single cell suspension of cells disassociated from sample that comprises cells from an individual, and culturing such cells in conditions suitable for culturing and maintaining human embryonic stem cells or in conditions comprising a serum free minimal medium supplemented with human basic fibroblast growth factor and human epidermal growth factor;

wherein multipotent adult stem cells are identified and isolated from said culture;

wherein said sample is selected from the group consisting of;

cells from follicular epithelium of a hair shaft;

a sample of liver tissue;

a sample of pancreatic tissue;

a sample of cardiac tissue; and

a sample comprising nerve cells.

24. (canceled)

26. A method of preparing an isolated population of human multipotent adult stem cells comprising the steps of:

a) forming a single cell suspension of cells disassociated from sample that comprises cells from an individual, and

b) culturing such cells in conditions suitable for culturing and maintaining human embryonic stem cells or in conditions comprising a serum free minimal medium supplemented with human basic fibroblast growth factor and human epidermal growth factor;

wherein multipotent adult stem cells are identified and isolated from said culture.

30. The method of claim 26 wherein said sample is selected from the group consisting of:

cells from follicular epithelium of a hair shaft;

a sample of liver tissue;

a sample of kidney, tissue;

a sample of pancreatic tissue;

a sample of cardiac tissue; and

a sample comprising nerve cells.

32. The method of claim 30 wherein said sample is cells from follicular epithelium of a hair shaft.

27-29. -29. (canceled)

31. (canceled)

33-41. -41. (canceled)