Hematopoietic stem cells and methods of treatment of neovascular eye diseases therewith
First Claim
1. An isolated, mammalian lineage negative hematopoietic stem cell population, which comprises hematopoietic stem cells and endothelial progenitor cells.
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Abstract
Isolated, mammalian, adult bone marrow-derived, lineage negative hematopoietic stem cell populations (Lin− HSCs) contain endothelial progenitor cells (EPCs) capable of rescuing retinal blood vessels and neuronal networks in the eye. Preferably at least about 20% of the cells in the isolated Lin− HSCs express the cell surface antigen CD31. The isolated Lin− HSC populations are useful for treatment of ocular vascular diseases. In a preferred embodiment, the Lin− HSCs are isolated by extracting bone marrow from an adult mammal; separating a plurality of monocytes from the bone marrow; labeling the monocytes with biotin-conjugated lineage panel antibodies to one or more lineage surface antigens; removing of monocytes that are positive for the lineage surface antigens from the plurality of monocytes, and recovering a Lin− HSC population containing EPCs. Isolated Lin− HSCs that have been transfected with therapeutically useful genes are also provided, and are useful for delivering genes to the eye for cell-based gene therapy. Methods of preparing isolated stem cell populations of the invention, and methods of treating ocular diseases and injury are also described.
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Citations
71 Claims
- 1. An isolated, mammalian lineage negative hematopoietic stem cell population, which comprises hematopoietic stem cells and endothelial progenitor cells.
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15. A method of isolating an adult bone marrow-derived, lineage negative hematopoietic stem population including endothelial progenitor cells comprising the steps of:
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(a) extracting bone marrow from an adult mammal;
(b) separating a plurality of monocytes from the bone marrow;
(c) labeling the monocytes with biotin-conjugated lineage panel antibodies to one or more lineage surface antigens selected from the group consisting of CD2, CD3, CD4, CD11, CD11a, Mac-1, CD14, CD16, CD19, CD24, CD33, CD36, CD38, CD45, Ly-6G, TER-119, CD45RA, CD56, CD64, CD68, CD86, CD66b, HLA-DR, and CD235a;
(d) removing monocytes that are positive for said one or more lineage surface antigens from the plurality of monocytes and recovering a population of lineage negative hematopoietic stem cells containing endothelial progenitor cells. - View Dependent Claims (16, 17, 18, 19, 20, 21)
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22. An isolated, mammalian, adult bone marrow-derived lineage negative hematopoietic stem cell population containing endothelial progenitor cells produced by:
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(a) extracting bone marrow from an adult mammal;
(b) separating a plurality of monocytes from the bone marrow;
(c) labeling the monocytes with biotin-conjugated lineage panel antibodies to one or more lineage surface antigens selected from the group consisting of CD2, CD3, CD4, CD11, CD11a, Mac-1, CD14, CD16, CD19, CD24, CD33, CD36, CD38, CD45, Ly-6G, TER-119, CD45RA, CD56, CD64, CD68, CD86, CD66b, HLA-DR, and CD235a;
(d) removing monocytes that are positive for said one or more lineage surface antigens from the plurality of monocytes and recovering a population of lineage negative hematopoietic stem cells containing endothelial progenitor cells;
wherein at least about 20% of the cells express the surface antigen CD31. - View Dependent Claims (23, 24, 25, 26, 27, 28, 29, 30)
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35. A method of treating an ocular disease in a mammal comprising isolating from the bone marrow of the mammal a lineage negative hematopoietic stem cell population that includes endothelial progenitor cells by:
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(a) extracting bone marrow from a mammal suffering from an ocular disease;
(b) separating a plurality of monocytes from the bone marrow;
(c) labeling the monocytes with biotin-conjugated lineage panel antibodies to one or more lineage surface antigens selected from the group consisting of CD2, CD3, CD4, CD11, CD11a, Mac-1, CD14, CD16, CD19, CD24, CD33, CD36, CD38, CD45, Ly-6G, TER-119, CD45RA, CD56, CD64, CD68, CD86, CD66b, HLA-DR, and CD235a;
(d) separating monocytes that are positive for said one or more lineage surface antigens from the plurality of monocytes and recovering a population of lineage negative hematopoietic stem cells containing endothelial progenitor cells;
and subsequently intravitreally injecting the isolated stem cells into an eye of the mammal in a number sufficient to ameliorate the effects of the disease. - View Dependent Claims (36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 56)
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61. A transfected lineage negative hematopoietic stem cell population prepared by:
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(a) extracting bone marrow from an adult mammal;
(b) separating a plurality of monocytes from the bone marrow;
(c) labeling the plurality of monocytes with biotin-conjugated lineage panel antibodies to CD2, CD3, CD4, CD11, CD11a, Mac-1, CD14, CD16, CD19, CD24, CD33, CD36, CD38, CD45, Ly-6G, TER-119, CD45RA, CD56, CD64, CD68, CD86, CD66b, HLA-DR, and CD235a;
(d) separating monocytes that are positive for said one or more lineage surface antigens from the plurality of monocytes and recovering a population of lineage negative hematopoietic stem cells containing endothelial progenitor cells; and
(e) transfecting the lineage negative hematopoietic stem cells recovered in step (d) with a polynucleotide that operably encodes a therapeutically useful peptide. - View Dependent Claims (62, 63, 65)
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Specification