Serum-free chemically defined cell culture medium
First Claim
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1. A serum-free chemically defined cell culture medium composition, the composition comprising:
- a serum free base medium comprising Dulbecco'"'"'s Modified Eagle Medium and MCDB 201 medium;
a regulated iron source;
an electron transport activator;
an antioxidant;
a steroid;
5-hydroxytryptamine;
a human serum albumin;
a phospholipid growth factor selected from lysophosphatidic acid and sphingosine 1-phosphate;
a cell growth factor;
human wingless/integrated 3a; and
R-spondin-1, said composition adjusted to a pH in a range of about 7.3 to about 7.5.
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Abstract
Embodiments of chemically defined cell culture media containing nutrients and growth factors free of any serum for culturing cells such as mesenchymal stem cells and methods of using embodiments of the cell culture medium for expanding cell populations such as mesenchymal stem cells while maintaining a pluripotent phenotype and methods of inducing chondrogenesis and osteogenesis of mesenchymal stem cells by admixing differentiation factors into embodiments of the cell culture medium.
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Citations
23 Claims
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1. A serum-free chemically defined cell culture medium composition, the composition comprising:
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a serum free base medium comprising Dulbecco'"'"'s Modified Eagle Medium and MCDB 201 medium; a regulated iron source; an electron transport activator; an antioxidant; a steroid; 5-hydroxytryptamine; a human serum albumin; a phospholipid growth factor selected from lysophosphatidic acid and sphingosine 1-phosphate; a cell growth factor; human wingless/integrated 3a; and R-spondin-1, said composition adjusted to a pH in a range of about 7.3 to about 7.5. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22)
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23. A serum-free chemically defined cell culture medium composition, the composition comprising:
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an amount of MCDB 201 medium of about 0.5 L/L; an amount of Dulbecco'"'"'s modified eagle medium of about 0.5 L/L; an amount of sodium bicarbonate of about 3.7 g/L; an amount of human serum albumin of about 0.25 g/L; an amount of transferrin of about 0.005 g/L; an amount of insulin of about 0.005 g/L; an amount of selenous acid of about 0.0000037 g/L; an amount of ascorbic acid-2phosphate of about 0.032205 g/L; an amount of 5-hydroxytryptamine of about 0.002127 g/L; an amount of dexamethasone of about 0.000003925 g/L; an amount of hydrocortisone of about 0.001 g/L; and an amount of α
-tocopherol acetate of about 0.0002 g/L;an amount of platelet-derived growth factor bb of about 10 ng/mL; an amount of human basic fibroblast growth factor of about 10 ng/mL; an amount of epiderman growth factor of about 20 ng/mL; an amount of insulin like growth factor-1 of about 5 ng/mL; an amount of lysophosphatidic acid of about 100 nM; an amount of sphingosine 1-phosphate of about 150 nM; an amount of human wingless/integrated 3a of about 20 ng/mL; and an amount of R-spondin-1 of about 40 ng/mL.
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Specification