STEM-LIKE CELLS IN BONE SARCOMAS
First Claim
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1. An isolated or purified population of pluripotent mammalian stem cells comprising:
- at least one embryonic stem cell marker on the surface of the stem cells, wherein the embryonic stem cell marker is Oct 3/4 or Nanog; and
,at least one stem cell marker comprising, activated Stat-3 CXCR4, CD 133, SCA-1, Tra-1-60, CD 44, CD 73, CD 90, CD 105 and Stro-1;
wherein,the purified population of mammalian pluripotent stem cells, is substantially free of cells that do not express the stem cell markers.
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Abstract
Isolation and purification of stem cells from within a bulk sarcoma tumor. These cells express the marker genes of pluripotent embryonic stem cells, Stat 3, Oct 3/4, and Nanog. A subset of these cells show the surface marker of mesenchymal stem cells Stro-1, as well as express attributes of mesodermal, ectodermal, and endodermal differentiation. The isolation, purification and characterization of these stem cells now provides the ideal target for the development of highly effective therapies against tumors.
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Citations
42 Claims
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1. An isolated or purified population of pluripotent mammalian stem cells comprising:
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at least one embryonic stem cell marker on the surface of the stem cells, wherein the embryonic stem cell marker is Oct 3/4 or Nanog; and
,at least one stem cell marker comprising, activated Stat-3 CXCR4, CD 133, SCA-1, Tra-1-60, CD 44, CD 73, CD 90, CD 105 and Stro-1;
wherein,the purified population of mammalian pluripotent stem cells, is substantially free of cells that do not express the stem cell markers. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A method of isolating stem cells from sarcomas comprising:
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isolating a sarcoma; dissociating cells from the sarcoma; culturing the cells on a non-adhesive substrate in anchorage independent serum free media; adding hormones and growth factors; adding methyl cellulose;
thereby,isolating stem cells. - View Dependent Claims (10, 11, 12, 13, 14)
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15. A method of treating tumors comprising:
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targeting stem cells expressing at least one marker selected from;
Oct 3/4, Nanog, activated Stat-3, CXCR4, CD 133, SCA-1, Tra-1-60, CD 44, CD 73, CD 90, CD 105 and Stro-1;inhibiting proliferation and/or lysing the stem cells; and
,inhibiting stem cell migration to a tumor and/or stem cell differentiation in a tumor;
wherein,growth and/or metastasis of a tumor is inhibited. - View Dependent Claims (16, 17, 18, 19, 20, 21, 22, 23, 24, 25)
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26. A method of treating a tumor comprising:
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isolating stem cells from a patient, whereby the stem cells are identified by Oct 3/4, Nanog, activated Stat-3, CXCR4, CD 133, SCA-1, Tra-1-60, CD 44, CD 73, CD 90, CD 105 and Stro-1; and
,isolating antigen specific lymphocytes from the patient; and
,co-culturing the stem cells and lymphocytes ex-vivo thereby priming the lymphocytes; and
,isolating activated lymphocytes from the co-culture; and
,re-infusing the activated lymphocytes into the patient;
thereby,activating an immune response to said stem cells and preventing self-renewal of the tumor. - View Dependent Claims (27, 28)
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29. A method of treating a tumor comprising:
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isolating stem cells from a patient, whereby the stem cells are identified by Oct 3/4, Nanog, activated Stat-3, CXCR4, CD 133, SCA-1, Tra-1-60, CD 44, CD 73, CD 90, CD 105 and Stro-1; and
,isolating antigen presenting cells from the patient; and
,co-culturing the stem cells and antigen presenting cells ex-vivo; and
,isolating the antigen presenting cells from the co-culture; and
,re-infusing the antigen presenting cells into the patient;
thereby,activating an immune response to said stem cells and preventing self-renewal of the tumor. - View Dependent Claims (30, 31)
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32. A method of treating cancer comprising:
administering at least one protein or peptide to a patient comprising Oct 3/4, Nanog, activated Stat-3, CXCR4, CD 133, SCA-1, Tra-1-60, CD 44, CD 73, CD 90, CD 105 and Stro-1;
thereby stimulating an immune response against stem cells expressing said proteins.
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33. A method of selectively treating cancer comprising:
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administering to a patient in need of treatment a therapeutically effective amount of a composition comprising antigen-activated dendritic cells, wherein dendritic cells are produced from proliferating cell cultures by a method comprising providing a tissue source comprising dendritic cell precursors; culturing the tissue source on a substrate and in culture medium to expand the number of dendritic cell precursors by allowing the dendritic cell precursors to proliferate;
wherein,said culture medium comprises GM-CSF and at least one other factor which inhibits the proliferation or maturation of non-dendritic cell precursors thereby increasing the proportion of dendritic cell precursors in the culture; and
,culturing the dendritic cell precursors for a period of time sufficient to allow them to mature into mature dendritic cells; wherein the dendritic cells are pulsed with stem cell derived antigen;
wherein,the dendritic cells process the stem-cell derived antigen to produce modified self-protein antigen, which is expressed by the dendritic cells and which activate an in vivo immune response which destroys stem cells expressing the stem-cell derived antigen. - View Dependent Claims (34, 35, 36, 37)
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38. A method of identifying candidate therapeutic compounds comprising:
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culturing stem cells expressing at least one marker selected from;
Oct 3/4, Nanog, activated Stat-3, CXCR4, CD 133, SCA-1, Tra-1-60, CD 44, CD 73, CD 90, CD 105 and Stro-1 with a candidate therapeutic agent;identifying candidate therapeutic agents which inhibit proliferation or growth, and/or lyse the stem cells and/or inhibit stem cell migration to a tumor and/or stem cell differentiation in a tumor;
wherein,growth and/or metastasis of a tumor is inhibited, and identifying a candidate therapeutic agent.
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39. The method of claim 39, wherein a candidate therapeutic agent comprises organic molecules, inorganic molecules, vaccines, antibodies, nucleic acid molecules, proteins, peptides and vectors expressing nucleic acid molecules.
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40. A method of treating cancer comprising:
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administering antibodies to stem cell biomarkers wherein the antibodies are specific for at least one marker selected from;
Oct 3/4, Nanog, activated Stat-3, CXCR4, CD 133, SCA-1, Tra-1-60, CD 44, CD 73, CD 90, CD 105 and Stro-1 and/or ligands thereof;inhibiting proliferation or growth, and/or lysing the stem cells and/or inhibiting stem cell migration to a tumor and/or stem cell differentiation in a tumor;
wherein, growth and/or metastasis of a tumor is inhibited, and treating cancer.
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41. A method of producing self-renewing pluripotent stem cell clones comprising:
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culturing sarcospheres; dissociating sarcospheres into single cells; culturing said single cells to near confluence and harvesting; reseeding harvested cells into suspension cultures; identifying self-renewing cells by formation of secondary spheres; thereby producing self-renewing pluripotent stem cell clones. - View Dependent Claims (42)
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Specification