Multipotent progenitor cell derived from adipose tissue

Abstract

Disclosed is a cell mass containing an adipose-tissue-derived multipotent progenitor cell. Also disclosed is a method for producing an adipose-tissue-derived multipotent progenitor cell from an adipose tissue, which comprises the steps of: (a) removing erythrocytes from an adipose-tissue-derived cell mass to produce a preadipose-tissue-derived multipotent progenitor cell mass; and (b) removing cells other than the adipose-tissue-derived multipotent progenitor cell from the preadipose-tissue-derived multipotent progenitor cell mass to produce the desired adipose-tissue-derived multipotent progenitor cell. Further disclosed is an adipose-tissue-derived multipotent progenitor cell produced by the method.

1 . A cell population containing an adipose tissue-derived multipotent progenitor cell, wherein the adipose tissue-derived multipotent progenitor cell expresses Islet-1 and at least one of Sca-1 and ABCG2. 
     
     
         2 - 10 . (canceled) 
     
     
         11 . A method for increasing purity of Islet-1 expressing adipose tissue-derived multipotent progenitor cell in a culture medium from an adipose tissue, comprising:
 (a) slicing and digesting adipose tissue to make an adipose cell extract of separated cells of adipose tissue-derived cell population;   (b) clarifying the separated cells from naturally occurring tissue and digestion products by centrifugation;   (c) selectively removing erythrocytes from the clarified adipose tissue-derived cell population by the density method;   (d) culturing in the presence of feeder cells and then washing the erythrocyte-free clarified adipose tissue-derived cell population; and   (e) selectively removing vascular endothelial cells from the cultured and washed erythrocyte-free clarified adipose tissue-derived cell population by treatment with EDTA, thereby forming a tissue culture enriched in Islet-1 expressing adipose tissue-derived multipotent progenitor cell population with respect to erythrocytes and respect to vascular endothelial cells, wherein the adipose tissue-derived multipotent progenitor cell expresses at least one of Sca-1 and ABCG2.   
     
     
         12 . The method according to  claim 11 , further comprising culturing the culture of Islet-1 expressing adipose tissue-derived multipotent progenitor cell in suspension medium to produce a hepatic lobule-like cell cluster. 
     
     
         13 . The method according to  claim 12 , wherein culturing the culture of Islet-1 expressing adipose tissue-derived multipotent progenitor cell in suspension medium comprises culturing for 3 to 4 weeks in 60% DMEM (low glucose), rhEGF, bFGF, HGF and OSM. 
     
     
         14 . The method according to  claim 13 , further comprising adding DMSO to the suspension culture medium. 
     
     
         15 . The method according to  claim 12 , wherein the hepatic lobule-like cluster comprises at least one hepatocyte. 
     
     
         16 . The method according to  claim 15 , further comprising adding DMSO to the suspension culture medium. 
     
     
         17 . The method according to  claim 11 , wherein the prepared culture lacks erythrocyte cells and vascular endothelial cells and contains feeder cells. 
     
     
         18 . The method according to  claim 11 , wherein the prepared tissue culture has 23 percent fibroblast cell contamination. 
     
     
         19 . A method for obtaining a hepatic lobule-like cell cluster, cardiac myoblasts or bone tissue from an Islet-1 expressing adipose tissue-derived multipotent progenitor cell from an adipose tissue, comprising:
 (a) slicing and digesting adipose tissue to make an adipose cell extract of separated cells of adipose tissue-derived cell population;   (b) clarifying the separated cells from naturally occurring tissue and digestion products by centrifugation;   (c) selectively removing erythrocytes from the adipose tissue-derived cell population by the density method, then culturing;   (d) culturing in the presence of feeder cells and then washing the erythrocyte-free clarified adipose tissue-derived cell population;   (e) selectively removing vascular endothelial cells from the cultured and washed erythrocyte-free clarified adipose tissue-derived cell population by a chelator; thereby forming a tissue culture enriched in Islet-1 expressing adipose tissue-derived multipotent progenitor cell population with respect to erythrocytes and respect to vascular endothelial cells, wherein the adipose tissue-derived multipotent progenitor cell expresses at least one of Sca-1 and ABCG2, and   (f) culturing said tissue culture enriched in Islet-1 expressing adipose tissue-derived multipotent progenitor cell population and obtaining a hepatic lobule-like cell cluster, cardiac myoblasts or bone tissue.