Adipose tissue-derived stromal cells and methods of isolating and using the same

Abstract

Adipose tissue-derived stromal cells and methods of isolating and using the same. In at least one embodiment of isolated adipose tissue-derived stromal cells of the present disclosure, the cells are isolated by performing adipose tissue resection or suction on a mammalian patient, dissecting tissue obtained from said tissue resection or suction and dissociating said tissue into a cell suspension, removing adipocytes from the cell suspension, culturing the adipocyte-depleted cell suspension in EGM-2-MV media, and isolating adipose tissue-derived stromal cells secreting vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and granulocyte-colony stimulating factor (G-CSF).

1 . An isolated adipose tissue-derived stromal cell. 
     
     
         2 . The isolated cell of  claim 1 , which expresses at least one characteristic of transformed cell selected from the group consisting of a cardiac cell, an endothelial cell, a smooth muscle cell, a dopaminergic neuronal cell, and a hepatic cell. 
     
     
         3 . 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). 
     
     
         4 . The isolated cell of  claim 3 , cultured on matrigel. 
     
     
         5 . The isolated cell of  claim 4 , cultured in DMEM and 10% fetal bovine serum which express alpha-actin. 
     
     
         6 . The isolated cell of  claim 1 , cultured in the presence of 5-azacytidine. 
     
     
         7 . 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. 
     
     
         8 . 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). 
     
     
         9 . The isolated cell of  claim 1 , which expresses CD34 and does not express VE-cadherin. 
     
     
         10 . The isolated cell of  claim 1 , wherein mechanical separation is performed on adipose tissue to dissociate the adipose tissue into a cell suspension containing the isolated adipose tissue-derived cell, the mechanical separation selected from the group consisting of mechanical agitation, sonic energy, thermal energy, and a combination of two or more of the same. 
     
     
         11 . The isolated cell of  claim 1 , wherein chemical separation is performed on adipose tissue to dissociate the adipose tissue into a cell suspension containing the isolated adipose tissue-derived cell, the chemical separation selected from the group consisting of collagenase separation, dispase separation, trypsin separation, and a combination of two or more of the same. 
     
     
         12 . A defined cell population comprising the cell of  claim 1 . 
     
     
         13 . The cell population of  claim 12 , wherein said cells comprise an exogenous nucleic acid encoding a protein of interest. 
     
     
         14 . The cell population of  claim 13 , 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. 
     
     
         15 . The isolated cell of  claim 1 , wherein the isolated cell is obtained from a mammalian patient and is subsequently administered back to the patient to treat the patient. 
     
     
         16 . The isolated cell of  claim 1 , wherein the isolated cell is initially harvested within a plurality of cells from a mammalian patient, wherein the harvesting is selected from the group consisting of liposuction and surgery. 
     
     
         17 . Adipose tissue-derived stromal cells, isolated by:
 performing adipose tissue resection or suction on a mammalian patient;   dissecting tissue obtained from said tissue resection or suction and dissociating said tissue into a cell suspension;   removing adipocytes from the cell suspension; and   isolating adipose tissue-derived stromal cells from the cell suspension with adipocytes removed.   
     
     
         18 . The adipose tissue-derived stromal cells of  claim 17 , wherein the step of removing adipocytes from the cell suspension further comprises the step of exposing the cell suspension to red cell lysis buffer. 
     
     
         19 . The adipose tissue-derived stromal cells of  claim 17 , wherein the adipose tissue-derived stromal cells are subsequently cultured in EGM-2-MV media to effectively multiply the number of the isolated adipose tissue-derived stromal cells. 
     
     
         20 . The adipose tissue-derived stromal cells of  claim 19 , wherein the adipose tissue-derived stromal cells secrete at least one factor selected from the group consisting of a vascular endothelial growth factor (VEGF), a hepatocyte growth factor (HGF), and a granulocyte-colony stimulating factor (G-CSF). 
     
     
         21 . Adipose tissue-derived stromal cells, isolated by:
 performing adipose tissue resection or suction on a mammalian patient;   dissecting tissue obtained from said tissue resection or suction and dissociating said tissue into a cell suspension;   removing adipocytes from the cell suspension;   culturing the adipocyte-depleted cell suspension in EGM-2-MV media; and   isolating adipose tissue-derived stromal cells secreting vascular endothelial growth factor (VEGF), hepatocyte growth factor (HGF), and granulocyte-colony stimulating factor (G-CSF).   
     
     
         22 . The adipose tissue-derived stromal cells of  claim 21 , wherein the isolated adipose tissue-derived stromal cells contain pluripotent cells capable of differentiating into another cell type.