Compositions and Methods Useful for Culturing Differentiable Cells

Abstract

The present invention relates to cell culture methods and compositions that are essentially serum-free and comprise a basal salt nutrient solution and an ErbB3 ligand.

1 - 34 . (canceled) 
     
     
         35 . Pluripotent cells in an in vitro suspension culture. 
     
     
         36 . The pluripotent cells of  claim 35 , wherein the pluripotent cells are substantially undifferentiated. 
     
     
         37 . The pluripotent cells of  claim 35 , wherein said pluripotent cells are human embryonic stem cells. 
     
     
         38 . The pluripotent cells of  claim 35 , wherein the suspension culture comprises an ErbB3 ligand. 
     
     
         39 . The pluripotent cells of  claim 35 , wherein said ErbB3 ligand is selected from the group consisting of Neuregulin-1, Heregulin-β (HRG-β), Heregulin-α (HRG-α), Neu differentiation factor (NDF), acetylcholine receptor-inducing activity (ARIA), glial growth factor 2 (GGF2), motor-neuron derived factor (SMDF), Neuregulin-2, Neuregulin-2β (NRG2-β), Epiregulin, Biregulin and functional fragments thereof. 
     
     
         40 . The pluripotent cells of  claim 35 , wherein the suspension culture comprises a TGF-β superfamily member. 
     
     
         41 . The pluripotent cells of  claim 35 , wherein said TGF-β family member is selected from the group consisting of Nodal, Activin A, Activin B, bone morphogenic protein-2 (BMP2), bone morphogenic protein-4 (BMP4) and functional fragments thereof. 
     
     
         42 . The pluripotent cells of  claim 35 , wherein the suspension culture comprises an insulin-like growth factor. 
     
     
         43 . The pluripotent cells of  claim 35 , wherein the suspension culture comprises a fibroblast growth factor. 
     
     
         44 . The pluripotent cells of  claim 35 , wherein said fibroblast growth factor is selected from the group consisting of FGF-2, FGF-7, FGF-10, FGF-22 and variants functional fragments thereof. 
     
     
         45 . The pluripotent cells of  claim 35 , wherein the suspension culture comprises a ROCK inhibitor. 
     
     
         46 . The pluripotent cells of  claim 35 , wherein said Rock inhibitor is Y27632. 
     
     
         47 . The pluripotent cells of  claim 35 , wherein the suspension culture does not comprise a TGF-β superfamily member. 
     
     
         48 . The pluripotent cells of  claim 35 , wherein the suspension culture does not comprise a fibroblast growth factor. 
     
     
         49 . The pluripotent cells of  claim 35 , wherein the suspension culture is serum free. 
     
     
         50 . The pluripotent cells of  claim 35 , wherein the suspension culture is matrix free. 
     
     
         51 . The pluripotent cells of  claim 35 , wherein the cells are maintained in suspension for at least two days. 
     
     
         52 . The pluripotent cells of  claim 35 , wherein the suspension culture comprises an erbB3 ligand and is serum free. 
     
     
         53 . An in vitro population of pluripotent cells in suspension culture. 
     
     
         54 . An in vitro suspension culture comprising pluripotent cells.