Cell culture method for mesenchymal stem cells

Abstract

The present disclosure relates to methods, cell culture medium and compositions that promote cell proliferation during fetal bovine serum free cell culture.

1 . A method of promoting stem cell proliferation in-vitro, the method comprising: culturing a population of mesenchymal lineage stem cells in a fetal bovine serum free cell culture medium comprising platelet derived growth factor (PDGF) and fibroblast growth factor 2 (FGF2), wherein the level of FGF2 is less than about 6 ng/ml. 
     
     
         2 . A fetal bovine serum free cell culture medium for promoting stem cell proliferation in-vitro comprising:
 a basal medium;   platelet derived growth factor (PDGF) fibroblast growth factor 2 (FGF2);   wherein the level of FGF2 is less than about 6 ng/ml.   
     
     
         3 . The method of  claim 1  or the culture medium of  claim 2 , wherein the level of FGF2 is between at least about 2 pg/ml and 40 pg/ml. 
     
     
         4 . The method of  claim 1  or the culture medium of  claim 2 , wherein the level of FGF2 is about 20 pg/ml. 
     
     
         5 . The method or culture medium of any one of  claims 1  to  4 , wherein the PDGF is PDGF-BB. 
     
     
         6 . The method or culture medium of  claim 5 , wherein the level of PDGF-BB is between at least about 3.0 ng/ml and about 120 ng/ml. 
     
     
         7 . The method or culture medium of  claim 5 , wherein the level of PDGF-BB is between about 9.0 ng/ml and about 60 ng/ml. 
     
     
         8 . The method or culture medium of  claim 5 , wherein the level of PDGF-BB is at least about 30 ng/ml. 
     
     
         9 . The method or culture medium of any one of  claims 1  to  8 , wherein the cell culture medium further comprises epidermal growth factor (EGF). 
     
     
         10 . The method or culture medium of  claim 9 , wherein the level of EGF is between about 0.08 ng/ml and about 7 ng/ml. 
     
     
         11 . The method or culture medium of  claim 9 , wherein the level of EGF is between about 0.4 ng/ml and about 1.6 ng/ml. 
     
     
         12 . The method or culture medium of  claim 9 , wherein the level of EGF is at least about 0.8 ng/ml. 
     
     
         13 . The method or culture medium of any one of  claims 1  to  12 , wherein the cell culture medium comprises alpha-minimal essential medium or fetal bovine serum free expansion medium. 
     
     
         14 . The method or culture medium of any one of  claims 1  to  13 , wherein the cell culture medium maintains the stem cells in an undifferentiated state. 
     
     
         15 . The method or culture medium of any one of  claims 1  to  14 , wherein the stem cell is a mesenchymal lineage precursor cell. 
     
     
         16 . The method of any one of  claims 1  to  15 , wherein the stem cell is a mesenchymal stem cell.