Isolation and purification of hematopoietic stem cells from post-liposuction lipoaspirates

Abstract

The present invention relates to a method of isolating hematopoietic stem cells from adipose tissue. The method yields a notably high number of CD34 + , ALDH br and/or ABCG2-expressing cells, comprising hematopoietic stem cells, permitting the use of the cells with no or minimal expansion.

What is claimed is: 
     
         1 . A method for isolating a CD34 +  cell, the method comprising:
 obtaining adipose tissue; 
 preparing a stromal vascular fraction (SVF) from the adipose tissue; 
 separating nonadherent cells in the SVF from the adherent cells; and 
 isolating a CD34 +  cell from the nonadherent cells, thereby isolating a CD34 +  cell. 
 
     
     
         2 . A composition comprising CD34 +  cells, the CD34 +  cells isolated according to the method of  claim 1 , wherein the CD34 +  cells are isolated in a ratio of at least about 1×10 5  CD34 +  cells isolated per about 1 milliliter of adipose tissue. 
     
     
         3 . The composition of  claim 2 , wherein the adipose tissue is human. 
     
     
         4 . A method of providing hematopoietic stem cells to a subject comprising,
 obtaining adipose tissue from a donor subject;   preparing a stromal vascular fraction (SVF) from the adipose tissue;   isolating nonadherent cells in the SVF;   isolating hematopoietic stem cells from the nonadherent cells; and   administering a therapeutically effective amount of the hematopoietic stem cells to a recipient subject in need thereof.   
     
     
         5 . The method of  claim 4 , wherein the donor subject and the recipient subject are human. 
     
     
         6 . The method of  claim 5 , wherein the donor subject and the recipient subject are the same human. 
     
     
         7 . The method of  claim 5 , wherein the donor subject and the recipient subject are not the same human. 
     
     
         8 . The method of  claim 4 , wherein the hematopoietic stem cells are CD34 +  and are isolated in the ratio of at least about 1×10 7  CD34 +  cells isolated per about 100 ml of adipose tissue. 
     
     
         9 . The method of  claim 6 , wherein the obtaining step comprises lipoaspiration. 
     
     
         10 . The method of  claim 4 , wherein the isolating step comprises isolating ALDH br SSC lo  cells or CD34 + CD38 − CD41 − SSC lo  cells. 
     
     
         11 . A composition of isolated cells comprising at least about 40% nonadherent, stromal vascular fraction (SVF) CD34 +  cells. 
     
     
         12 . The composition of claim  25 , wherein the isolated cells comprise at least about 20% CD34 + CD45 −  cells. 
     
     
         13 . The composition of  claim 11 , wherein the stromal vascular fraction is obtained from human adipose tissue. 
     
     
         14 . The composition of  claim 11 , wherein the CD34 +  cells comprise CD34 + CD38 − CD41 − ALDH br  cells. 
     
     
         15 . The composition of  claim 11 , wherein the CD34 +  cells comprise CD34 + CD38 − CD41 − SSC lo  cells. 
     
     
         16 . The composition of  claim 11 , wherein the CD34 +  cells comprise colony-forming unit granulocyte macrophages (CFU-GM), burst-forming unit erythrocytes (BFU-E) and colony-forming unit granulocyte erythrocyte macrophage megakaryocytes (CFU-GEMM). 
     
     
         17 . The composition of  claim 11 , wherein the cells are genetically modified. 
     
     
         18 . A composition of isolated cells comprising at least about 10% nonadherent, stromal vascular fraction (SVF) ALDH br  cells. 
     
     
         19 . The composition of  claim 18 , comprising at least about 15% nonadherent SVF ALDH br  cells. 
     
     
         20 . The composition of  claim 18 , wherein the ALDH br  cells are ALDH br SSC lo  cells. 
     
     
         21 . The composition of  claim 18 , wherein the SVF is obtained from human adipose tissue. 
     
     
         22 . A pharmaceutical composition comprising the composition according to  claim 2  and a pharmaceutically acceptable carrier. 
     
     
         23 . A pharmaceutical composition comprising the composition according to  claim 11  and a pharmaceutically acceptable carrier. 
     
     
         24 . A pharmaceutical composition comprising the composition according to  claim 18  and a pharmaceutically acceptable carrier.