Identification and isolation of multipotent cells from non-osteochondral mesenchymal tissue

Abstract

Methods for the identification and isolation of multipotent cells from non-osteochondral mesenchymal tissue. Specifically, this invention relates to an adult multipotent cell or a cell population or composition comprising said cell, isolated from non-osteochondral mesenchymal tissue, characterized in that the cell 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: CD11b, CD14, CD15, CD16, CD31, CD34, CD45, CD49f, CD102, CD104, CD106 and CD133.

1 - 20 . (canceled)  
   
   
       21 . 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.  
   
   
       22 . An isolated multipotent cell of  claim 21 , 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, and eliminating cells that show no adhesion.  
   
   
       23 . An isolated multipotent cell according to  claim 21 , wherein the non-osteochondral mesenchymal tissue is connective tissue or adipose tissue.  
   
   
       24 . An isolated multipotent adult cell according to  claim 21 , wherein the cell is genetically modified.  
   
   
       25 . A differentiated cell that expresses at least one characteristic of a specialized cell, wherein the cell is derived from an isolated multipotent adult stem cell according to  claim 21 .  
   
   
       26 . The differentiated cell according to  claim 25 , 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.  
   
   
       27 . An isolated cell population that comprises cells according to  claim 21 .  
   
   
       28 . The isolated cell population according to  claim 27 , wherein the cells are nearly homogenous.  
   
   
       29 . A method for identifying a population of multipotent adult cells, wherein the population consists of an isolated cell according to  claim 21 , the method comprising: (a) incubating the cells with labeled specific binding compounds for one or more characteristic markers for the population; and (b) detecting the presence or absence of binding by the cells to these specific binding compounds.  
   
   
       30 . The method according to  claim 29 , wherein the specific binding compound is an antibody.  
   
   
       31 . A method for isolating a population of multipotent adult cells, wherein the cells have a profile of marker expression which is that the cells express surface markers CD9<+>, CD10<+>, CD13<+>, CD29<+>, CD44<+>, CD49A<+>, CD51<+>, CD54<+>, CD55<+>, CD58<+>, CD59<+>, CD90<+> and CD105<+>; and lack expression of surface markers CD11b, CD14, CD15, CD16, CD31, CD34, CD45, CD49f, CD102, CD104, CD106 and CD133, the method comprising: 
 (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 labeled compound that binds specifically to one or more of the surface makers; and    (d) selecting those cells that have the profile of expression of markers.    
   
   
       32 . The method according to  claim 31 , wherein a negative selection is performed, whereby cells are excluded that show binding to labeled compounds that bind specifically to a marker selected from the group consisting of CD11b, CD14, CD15, CD16, CD31, CD34, CD45, CD49f, CD102, CD104, CD106, CD133.  
   
   
       33 . The method according to  claim 32 , wherein a positive selection is performed, whereby cells are selected that bind to labeled 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.  
   
   
       34 . The method according to  claim 31 , wherein the labeled compound of specific binding is an antibody.  
   
   
       35 . A pharmaceutical composition that comprises a population of cells according to  claim 21  and a pharmaceutically acceptable carrier.  
   
   
       36 . A therapeutic method comprising administering a pharmaceutical composition of  claim 35  to a patient in need thereof.  
   
   
       37 . A therapeutic method according to  claim 36 , wherein the method is for tissue repair or regeneration.  
   
   
       38 . A method for assessing in vitro cell response to a biological or pharmacological agent, or to a combinatorial library the agents, the method comprising: 
 (a) isolating a cell population according to  claim 28 ,    (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.    
   
   
       39 . The method according to  claim 38 , wherein prior to step (c) the cells are allowed to differentiate into a specific type of cells.