Culture media for multipotent stem cells
A culture medium for human adipose tissue derived mesenchymal stem cells includes a keratinocyte-SFM basal medium, L-Cysteine, FGF and SDF-1 is disclosed. The culture medium is effective for growing mesenchymal stem cells at a high proliferation rate while maintaining the purity, characterization and undifferentiated state of the cells.
1. A stem cell growth medium comprising:
(a) serum about 1% to about 10% by volume;
(b) fibroblast growth factor 1 to about 10 ng/ml;
(c) epidermal growth factor 1 to about 10 ng/ml;
(d) hydrocortisone about 10 to about 100 ng/ml;
(e) calcium chloride 0.01 to about 0.1 mM;
(f) insulin about 0.5 to about 5 mg/ml;
(g) bovine pituitary extract about 10 to about 100 μg/ml;
(h) L-cysteine or glutathione about 0.5 to about 5 mM;
(i) selenium about 0.1 to about 1 μg/ml;
(j) stromal-derived factor 1 to about 10 ng/ml;
(k) sodium pyruvate 2 to about 20 mg/ml;
(l) transferrin 0.1 to about 1 mg/ml; and
(m) serum free medium balance to 100% volume.
2. The stem cell growth medium of claim 1 , wherein the serum is human serum.
3. The stem cell growth medium of claim 1 , wherein the serum is fetal bovine serum.
4. The stem cell growth medium of claim 1 , wherein at least one of the FGF or the EGF is recombinant.
5. The stem cell growth medium of claim 1 , wherein the SDF is recombinant SDF.
6. The stem cell growth medium of claim 1 , wherein at least one of the insulin or the transferrin is recombinant.
7. The stem cell growth medium of claim 1 , wherein said medium contains L-cysteine.
8. The stem cell growth medium of claim 1 , wherein said medium contains glutathione.
9. The stem cell growth medium of claim 1 , wherein the serum free medium is Keratinocyte-SFM, Defined Keratinocyte-SFM, MCDB 153, Keratinocyte Growth Medium, DMEM/F12, Ham's F10, Ham's F12 or RPMI 1640.
10. A method of culturing a cell, the method comprising culturing the cell in the stem cell growth medium of claim 1 .
11. The method of claim 10 , wherein said cell is a multipotent stem cell.
12. The method of claim 11 , wherein said multipotent stem cell is derived from adipose tissue.
13. The method of claim 11 , wherein said multipotent stem cell undergoes clonal expansion.
14. The method of claim 11 , wherein said multipotent stem cell is growing attached to a plastic material, showing spindle-shaped morphological features.
15. The method of claim 11 , wherein said multipotent stem cell is induced to undergo adipogenic differentiation.
16. The method of claim 11 , wherein said multipotent stem cell is induced to undergo chondrogenic differentiation.
17. The method of claim 11 , wherein said multipotent stem cell is induced to undergo osteogenic differentiation.
18. The method of claim 11 , wherein said multipotent stem cell expresses immunologically detectable surface expression of responses to CD73, CD90 or CD105, and/or expresses negative immunologically detectable surface expression of CD34, CD45 or HLA-DR.
19. A kit comprising ingredients for preparation of the stem cell growth medium according to claim 1 .