Identification and Isolation of Multipotent Cells From Non-Osteochondral Mesenchymal Tissue

Abstract

Identification and isolation of multipotent cells from non-osteochondral mesenchymal tissue. This invention relates to the identification and isolation of multipotent cells from non-osteochondral mesenchymal tissue. Specifically, it relates to an adult multipotent cell or a cell population or composition comprising said cell, isolated from non-osteochondral mesenchymal tissue, characterized in that it is positive for the following markers: CD9, CD10, CD13, CD29, CD44, CD49A, CD51, CD54, CD55, CD58, CD59, CD90 and CD105 and because it lacks expression of the following markers: CD11 b, CD14, CD15, CD16, CD31, CD34, CD45, CD49 f, CD102, CD104, CD106 and CD133.

1 . An isolated multipotent adult cell, which (a) is isolated from non-osteochondral mesenchymal tissue, (b) expresses CD9<+>, CD10<+>, CD13<+>, CD29<+>, CD44<+>, CD49A<+>, CD51<+>, CD54<+>, CD55<+>, CD58<+>, CD59<+>, CD90<+> and CD105<+>; and (c) lacks expression of CD11b, CD14, CD15, CD16, CD31, CD34, CD45, CD49f, CD102, CD104, CD106 and CD133.  
   
   
       2 . An isolated multipotent adult cell according to  claim 1 , which is isolated by a method that comprises: 
 (a) collecting a non-osteochondral mesenchymal tissue;    (b) obtaining a cell suspension by enzymatic digestion;    (c) sedimenting and resuspending the cells in a culture medium; and    (d) culturing the cells on a solid surface, and eliminating cells that show no adhesion to said solid surface.    
   
   
       3 . The isolated multipotent adult cell according to  claim 1 , wherein the non-osteochondral mesenchymal tissue is connective tissue.  
   
   
       4 . The isolated multipotent adult cell according to  claim 3 , wherein said connective tissue is adipose tissue.  
   
   
       5 . The isolated multipotent adult cell according to  claim 1 , wherein the cell is a genetically modified cell.  
   
   
       6 . A cell that expresses at least one characteristic of a specialized cell, wherein the cell is derived from an isolated multipotent adult cell according to  claim 1 .  
   
   
       7 . The cell according to  claim 6 , wherein the at least one characteristic is that of a cell selected from the group consisting of an epithelial cell, an endothelial cell, an adipocyte, a myocyte, a chondrocyte, an osteocyte, a neuron, an astrocyte, an oligodendrocyte, a hepatocyte, a cardiomyocyte, and a pancreatic cell.  
   
   
       8 . An isolated cell population that comprises cells according to  claim 1 .  
   
   
       9 . The isolated cell population according to  claim 8 , wherein said cell population is nearly homogeneous.  
   
   
       10 . A method for identifying a population of multipotent adult cells, wherein said population comprises isolated cells according to  claim 1 , the method comprising: (a) incubating the cells with labelled specific binding compounds for one or more characteristic markers for said population; and (b) detecting the presence or absence of binding by the cells to these specific binding compounds.  
   
   
       11 . The method according to  claim 10 , wherein the specific binding compound is an antibody.  
   
   
       12 . A method for isolating a population of multipotent adult cells according to  claim 1 , which comprises: 
 (a) collecting a non-osteochondral mesenchymal tissue;    (b) obtaining a cell suspension from the tissue by enzymatic digestion;    (c) incubating the cell suspension with a labelled compound that binds specifically to one or more of the surface makers of said multipotent adult cells; and    (d) selecting those cells that have the desired profile of expression of markers.    
   
   
       13 . The method according to  claim 12 , wherein a negative selection is performed, whereby cells are excluded that show binding to labelled compounds that bind specifically to a marker selected from the group consisting of CD11b, CD14, CD15, CD16, CD31, CD34, CD45, CD49f, CD102, CD104, CD106 and CD133.  
   
   
       14 . The method according to  claim 12 , wherein a positive selection is performed, whereby cells are selected that bind to labelled compounds that bind specifically to a marker selected from the group consisting of CD9, CD10, CD13, CD29, CD44, CD49a, CD51, CD54, CD55, CD58, CD59, CD90, and CD105.  
   
   
       15 . The method according to  claim 12 , wherein the labelled compound of specific binding is an antibody.  
   
   
       16 . A substantially homogeneous cell composition which comprises a multipotent adult cell according to  claim 1 .  
   
   
       17 . A pharmaceutical composition that comprises a multipotent adult cell according to  claim 1 , and a pharmaceutically acceptable carrier.  
   
   
       18 . An isolated multipotent adult cell according to  claim 1  for use selected from the group consisting of: in therapy; and in the repair and regeneration of tissues.  
   
   
       19 . (canceled)  
   
   
       20 . (canceled)  
   
   
       21 . A therapeutic method comprising administering a pharmaceutical composition according to  claim 17  to a patient in need thereof.  
   
   
       22 . Method according to  claim 21 , wherein said therapeutic method is for tissue repair or regeneration.  
   
   
       23 . A method for assessing in vitro cell response to a biological or pharmacological agent, or to a combinatorial library of said agents, the method comprising: 
 a) isolating a cell population according to  claim 8 ,    b) expanding the cell population via culturing, and    c) applying a biological agent or pharmacological agent or a combinatorial library of said agents to the cell population, and assessing effects of said agents on the cultured cells.    
   
   
       24 . Method according to  claim 23 , wherein prior to step (c) the cells are allowed to differentiate into a specific type of cells.