CELL CULTURE METHOD FOR MESENCHYMAL STEM CELLS
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.