Method of Deriving Mesenchymal Stem Cells

Abstract

We describe a method of obtaining a cell culture, the method comprising providing a cell obtained by dispersing a human embryonic stem cell (hESC) colony, or a descendent thereof, and propagating the cell in the absence of a feeder cell layer in a serum free medium comprising FGF2 and optionally PDGF AB. Preferably, the human embryonic stem cell (hESC) colony is dispersed with a dispersing agent which is trypsin.

1 - 54 . (canceled) 
     
     
         55 . A cell line obtained by providing a cell obtained by dispersing an embryonic cell colony or a descendent thereof and propagating the cell in the absence of co-culture in a serum free medium comprising FGF2. 
     
     
         56 . A method of conditioning a cell culture medium, the method comprising obtaining a cell by dispersing an embryonic cell colony or a descendent thereof, propagating the cell in the absence of co-culture in a serum free medium comprising FGF2, and culturing a mesenchymal stem cell, mesenchymal stem cell line or differentiated mesenchymal stem cell in a cell culture medium. 
     
     
         57 . A conditioned medium obtained by obtaining a cell by dispersing an embryonic cell colony or a descendent thereof, propagating the cell in the absence of co-culture in a serum free medium comprising FGF2, and culturing a mesenchymal stem cell, mesenchymal stem cell line or differentiated mesenchymal stem cell in a cell culture medium. 
     
     
         58 . The conditioned medium according to  claim 57 , wherein said medium is used to treat a disease in an individual. 
     
     
         59 . The cell line according to  claim 55 , wherein said embryonic cell colony is a human embryonic cell colony. 
     
     
         60 . The cell line according to  claim 55 , wherein said embryonic cell colony is dispersed with trypsin. 
     
     
         61 . The cell line according to  claim 55 , having a cell surface marker selected from the group consisting of CD105 and CD73. 
     
     
         62 . A cell line obtained by providing a cell obtained by dispersing an embryonic cell colony or a descendent thereof, propagating the cell in the absence of co-culture in a serum free medium containing FGF2, and selecting a mesenchymal stem cell from the propagated cells based on expression of a cell surface marker, and deriving a cell line from the cell selected therefrom. 
     
     
         63 . The cell line according to  claim 62 , wherein said embryonic cell colony is a human embryonic cell colony. 
     
     
         64 . The cell line according to  claim 62 , wherein said embryonic cell colony is dispersed with trypsin. 
     
     
         65 . The cell line according to  claim 62 , having a cell surface marker selected from the group consisting of CD105 and CD73. 
     
     
         66 . The method according to  claim 56 , further comprising the step of isolating the cell culture medium.