Adipose stromal stem cells for tissue and vascular modification

Abstract

The present invention provides isolated adipose derived stromal cells and methods of use thereof.

1 . An isolated adipose tissue derived stromal cell.  
   
   
       2 . The isolated cell of  claim 1 , which expresses at least one characteristic of a cardiac cell.  
   
   
       3 . The isolated cell of  claim 1 , which expresses at least one characteristic of an endothelial cell.  
   
   
       4 . The isolated cell of  claim 1 , which expresses at least one characteristic of a smooth muscle cell.  
   
   
       5 . The isolated cell of  claim 1 , which expresses at least one characteristic of a dopaminergic neuronal cell.  
   
   
       6 . The isolated cell of  claim 1 , which expresses at least one characteristic of a hepatic cell.  
   
   
       7 . The isolated cell of  claim 1  which secretes at least one factor selected from the group consisting of vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), granulocyte-colony stimulating factor (G-CSF), and basic fibroblast growth factor (bFGF).  
   
   
       8 . The isolated cell of  claim 3 , cultured on matrigel.  
   
   
       9 . The isolated cell of  claim 4 , cultured in DMEM and 10% fetal bovine serum which express alpha-actin.  
   
   
       10 . The isolated cell of  claim 2 , cultured in the presence of 5-azacytidine.  
   
   
       11 . The isolated cell of  claim 1 , which secretes a factor selected from the group consisting of pro-angiogenic factors, anti-apoptotic factors, vasculoprotective factors, and cardioprotective factors.  
   
   
       12 . The isolated cell of  claim 1 , initially cultured in EGM-2-MV media followed by subsequent culture in EBM-2 media, said cell secreting vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and granulocyte-macrophage colony stimulating factor (GM-CSF) and granulocyte-colony stimulating factor (G-CSF).  
   
   
       13 . The isolated cell of  claim 1 , which expresses CD34 and does not express VE-cadherin.  
   
   
       14 . A defined cell population comprising the cell of  claim 1 .  
   
   
       15 . The cell population of  claim 10 , wherein said cells comprise an exogenous nucleic acid encoding a protein of interest.  
   
   
       16 . The cell population of  claim 11 , wherein said exogenous nucleic acid is selected from the group of nucleic acids encoding VEGF, bFGF, IGF1, IGF2, HGF, cardiotrophin, myotrophin, nitric oxide synthase 1, nitric oxide synthase 2, nitric oxide synthase  3 , tumor necrosis factor alpha, tumor necrosis factor beta, fibroblast growth factor, pleotrophin, endothelin, and angiopoietin.  
   
   
       17 . A method for promoting angiogenesis in a mammal, comprising administration of therapeutic quantity of adipose-derived stromal cells to said mammal, such that therapeutic angiogenesis occurs.  
   
   
       18 . The method of  claim 17 , wherein said adipose-derived stromal cells are administered via a method selected from the group consisting of retrograde coronary venous delivery, retrograde delivery into other tissues, direct microinjection into target tissue, intra-arterial infusion via a catheter, intra-coronary infusion via a catheter, and systemic intravenous administration.  
   
   
       19 . The method of  claim 17 , wherein angiogenesis is promoted in cardiac tissue and said cells are administered via retrograde coronary venous delivery.  
   
   
       20 . The method of  claim 19 , wherein retrograde coronary venous delivery is conducted at pressures ranging from 30-400 mm Hg.  
   
   
       21 . The method of  claim 20 , wherein cells so delivered enter the interstitium.  
   
   
       22 . The method of  claim 17 , wherein said cells are administered in a biologically compatible medium.  
   
   
       23 . The method of  claim 22 , wherein said medium further comprises a viscosity-increasing carrier.  
   
   
       24 . The method of  claim 17 , wherein angiogenesis is promoted in ischemic tissue.  
   
   
       25 . A method for isolating adipose derived stromal cells from a human patient comprising: 
 a) performing adipose tissue resection or suction on said patient;    b) dissecting tissue obtained from said tissue resection or suction and dissociating said tissue into a cell suspension;    c) removing adipocytes from said cell suspension;    d) optionally exposing the cell suspension of step d) to red cell lysis buffer; and    e) isolating adipose derived stromal cells.    
   
   
       26 . The method of  claim 25 , further comprising culturing the cells of step e) in a culture medium of interest.  
   
   
       27 . The method of  claim 26 , wherein the cells of step e) are cultured in DMEM+10% fetal bovine serum and express alpha-actin.  
   
   
       28 . The method of  claim 26 , wherein the cells of step e) are initially cultured in EGM-2-MV media followed by subsequent culture in EBM-2 media, said cells secreting vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and granulocyte-colony stimulating factor (G-CSF).  
   
   
       29 . The method of  claim 25 , further comprising introduction of an exogenous nucleic acid molecule encoding a protein of interest into the cells of step e).  
   
   
       30 . The method of  claim 26 , wherein said cells are exposed to a receptor ligand cocktail, said ligands being selected from the group consisting of at least one of VEGF, LIF, bFGF, IGF1, IGF2, HGF, cardiotrophin, myotrophin, nitric oxide synthase 1, nitric oxide synthase 2, nitric oxide synthase 3, tumor necrosis factor alpha, tumor necrosis factor beta, fibroblast growth factor, pleotrophin, endothelin, and angiopoietin.  
   
   
       31 . A method for promoting cardiomyogenesis in a mammal, comprising administration of therapeutic quantity of adipose-derived stromal cells to said mammal, such that therapeutic cardiomyogenesis occurs.  
   
   
       32 . The method of  claim 31 , wherein said adipose-derived stromal cells are administered via a method selected from the group consisting of retrograde coronary venous delivery, retrograde delivery into other tissues, direct microinjection into target tissue, intra-arterial infusion via a catheter, intra-coronary infusion via a catheter, and systemic intravenous administration.  
   
   
       33 . The method of  claim 31 , wherein said cells are administered via retrograde coronary venous delivery.  
   
   
       34 . The method of  claim 33 , wherein retrograde coronary venous delivery is conducted at pressures ranging from 30-400 mm Hg.  
   
   
       35 . The method of  claim 34 , wherein cells so delivered enter the interstitium.  
   
   
       36 . The method of  claim 31 , wherein said cells are administered in a biologically compatible medium.  
   
   
       37 . The method of  claim 31 , wherein said medium further comprises a viscosity-increasing carrier.  
   
   
       38 . The method of  claim 31 , wherein cardiomyogenesis is promoted in ischemic cardiac tissue.  
   
   
       39 . The method of  claim 31 , wherein said cells further comprise an exogenous nucleic acid encoding a protein of interest.  
   
   
       40 . A method for promoting cardiomyogenesis in a mammal, comprising retrograde coronary venous delivery of a therapeutic quantity of adipose-derived stromal cells immediately following isolation of said cells, to said mammal, such that therapeutic cardiomyogenesis occurs.  
   
   
       41 . A method for repopulating bone marrow comprising administration of therapeutic quantity of adipose-derived stromal cells to said mammal, said stem cells differentiating into cells of hematopoietic lineage, thereby repopulating bone marrow.  
   
   
       42 . The method of  claim 41 , wherein said adipose-derived stromal cells are administered via a method selected from the group consisting of retrograde coronary venous delivery, retrograde delivery into other tissues, direct microinjection into target tissue, intra-arterial infusion via a catheter, intra-coronary infusion via a catheter, and systemic intravenous administration.  
   
   
       43 . The method of  claim 41 , wherein said cells are administered via retrograde coronary venous delivery.  
   
   
       44 . The method of  claim 43 , wherein retrograde coronary venous delivery is conducted at pressures ranging from 30-400 mm Hg.  
   
   
       45 . The method of  claim 41 , wherein said cells are administered in a biologically compatible medium.  
   
   
       46 . The method of  claim 42 , wherein said cells further comprise an exogenous nucleic acid encoding a protein of interest.  
   
   
       47 . A method for promoting hematopoiesis in a mammal, comprising administration of a therapeutic quantity of adipose-derived stromal cells to said mammal, such that therapeutic hematopoiesis occurs.