Hematopoietic stem cells and methods of treatment of neovascular eye diseases therewith

Abstract

Isolated, mammalian, bone marrow-derived, hematopoietic stem cell populations contain endothelial progenitor cells (EPC) capable of forming retinal blood vessels. At least about 50% of the cells in the isolated cell population express CD31 and c-kit. Up to about 8% of the cells can express Sca-1, and up to about 4% of the cells can express Flk-1/KDR. The cells can incorporate into the vasculature of the retina when intravitreally injected into the eye, and can thereby stabilize or rebuilt diseased or abnormal retinal blood vessels.

1 . An isolated, mammalian, bone marrow-derived hematopoietic stem cell population comprising endothelial progenitor cells in which at least about 50% of the total cells of the cell population express CD31 and c-kit, and not more than about 1 percent of the total cells of the cell population express Tie-2.  
     
     
         2 . The isolated cell population of  claim 1  wherein at least 75% of the total cells of the cell population express CD31.  
     
     
         3 . The isolated cell population of  claim 1  wherein at least 65% of the total cells of the cell population express c-kit.  
     
     
         4 . The isolated cell population of  claim 1  wherein at least 80% of the total cells of the cell population express CD31 and at least 70% of the total cells of the cell population express c-kit.  
     
     
         5 . The isolated cell population of  claim 1  wherein the cell population comprises murine cells.  
     
     
         6 . The isolated cell population of  claim 1  wherein the cell population comprises human cells.  
     
     
         7 . The isolated cell population of  claim 1  wherein up to about 8% of the total cells of the cell population express Sca-1 and up to about 4% of the total cells of the cell population express Flk-1/KDR.  
     
     
         8 . The isolated cell population of  claim 1  wherein about 50% to about 85% of the total cells of the cell population express CD31, about 70% to about 75% of the total cells of the cell population express c-kit, about 4% to about 8% of the total cells of the cell population express Sca-1, and about 2% to about 4% of the total cells of the cell population express Flk-1/KDR.  
     
     
         9 . The isolated cell population of  claim 1  further including a cell culture medium.  
     
     
         10 . The isolated cell population of  claim 1  wherein the cell population is obtained from mammalian bone marrow.  
     
     
         11 . The isolated cell population of  claim 1  wherein the cell population is obtained from human bone marrow.  
     
     
         12 . A method of isolating bone marrow-derived, hematopoietic stem cells including endothelial progenitor cells comprising the steps of: 
 (a) extracting bone marrow from a mammal;    (b) separating a plurality of monocytes from the bone marrow;    (c) labeling the plurality of monocytes with biotin conjugated lineage panel antibodies to CD45, CD3, Ly-6G, CD11 and TER-119; and    (d) removing monocytes that were lineage positive for CD45, CD3, Ly-6G, CD11 and TER-119 from the plurality of monocytes to provide a population of hematopoietic stem cells including endothelial progenitor cells.    
     
     
         13 . The method of  claim 12  wherein the mammal is an adult mammal.  
     
     
         14 . The method of  claim 12  wherein the mammal is a mouse.  
     
     
         15 . The method of  claim 12  wherein the mammal is a human.  
     
     
         16 . The method of  claim 12  wherein at least about 50% of the total cells of the cell population express CD31 and c-kit.  
     
     
         17 . The method of  claim 12  wherein the population of hematopoietic stem cells includes endothelial progenitor cells capable of targeting glial enriched regions of injured adult retinas.  
     
     
         18 . An isolated, mammalian, bone marrow-derived hematopoietic stem cell population produced by the method of  claim 12 .  
     
     
         19 . The isolated cell population of  claim 18  wherein at least about 50% of the total cells of the cell population express CD31 and c-kit, and not more than about 1% of the total cells of the cell population express Tie-2.  
     
     
         20 . The isolated cell population of  claim 18  wherein the cell population comprises murine cells.  
     
     
         21 . The isolated cell population of  claim 18  wherein the cell population comprises human cells.  
     
     
         22 . A method of enhancing retinal neovacularization in a mammal comprising intravitreally injecting cells from the cell population of  claim 1  into the eye of a mammal in need of retinal neovascularization, wherein the cell population is obtained from bone marrow of the same species of mammal as the species into whose eye the cells are injected.  
     
     
         23 . The method of  claim 22  wherein the mammal is a mouse.  
     
     
         24 . The method of  claim 22  wherein the mammal is a human.  
     
     
         25 . A method of treating an ocular disease in a mammal comprising isolating from the bone marrow of the mammal a hematopoietic stem cell population that includes endothelial progenitor cells and intravitreally injecting cells from the cell population into an eye of the mammal in an amount sufficient to arrest the disease, wherein at least about 50% of the total cells of the cell population express CD31 and c-kit, and not more than about 1 percent of the total cells of the cell population express Tie-2.  
     
     
         26 . The method of  claim 25  wherein the amount of cells injected is effective for repairing retinal damage in the eye.  
     
     
         27 . The method of  claim 25  wherein the amount of cells injected is effective for stabilizing retinal neovasculature of the eye.  
     
     
         28 . The method of  claim 25  wherein the amount of cells injected is effective for maturing retinal neovasculature of the eye.  
     
     
         29 . The method of  claim 25  wherein the cell population is isolated by: 
 (a) extracting bone marrow from the mammal;    (b) separating a plurality of monocytes from the bone marrow;    (c) labeling the plurality of monocytes with biotin conjugated lineage panel antibodies to CD45, CD3, Ly-6G, CD11 and TER-119; and    (d) removing monocytes that were lineage positive for CD45, CD3, Ly-6G, CD11 and TER-119 from the plurality of monocytes to provide a population of hematopoietic stem cells including endothelial progenitor cells.    
     
     
         30 . The method of  claim 25  wherein the mammal is a human.  
     
     
         31 . The method of  claim 25  wherein the disease is selected from the group consisting of a retinal degenerative disease, a retinal vascular degenerative disease, an ischemic retinopathy, a vascular hemorrhage, a vascular leakage, a choroidopathy, age related macular degeneration, diabetic retinopathy, presumed ocular histoplasmosis, retinopathy of prematurity, sickle cell anemia, and retinitis pigmentosa.  
     
     
         32 . A method of inducing neurotrophic rescue in a retina of a mammal suffering from a retinal degenerative disease comprising intravitreally injecting a neurotrophic rescue-inducing amount of cells from the cell population of  claim 1  into a diseased eye of the mammal.  
     
     
         33 . The method of  claim 32  wherein the cell population is isolated by: 
 (a) extracting bone marrow from the mammal;    (b) separating a plurality of monocytes from the bone marrow;    (c) labeling the plurality of monocytes with biotin conjugated lineage panel antibodies to CD45, CD3, Ly-6G, CD11 and TER-119; and    (d) removing monocytes that were lineage positive for CD45, CD3, Ly-6G, CD11 and TER-119 from the plurality of monocytes to provide a population of hematopoietic stem cells including endothelial progenitor cells.    
     
     
         34 . The method of  claim 32  wherein the mammal is a human.