UMBILICAL CORD LINING STEM CELLS AND METHODS AND MATERIAL FOR ISOLATING AND CULTURING SAME
First Claim
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1. A method for isolating umbilical cord lining stem cells (ULSCs) from an umbilical cord, said method comprisinga) obtaining the lining of an umbilical cord, wherein said lining is substantially free of blood, venous tissue, and arterial tissue;
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Abstract
Human umbilical cord lining stem cells that are capable of differentiating into cells of the mesodermal lineage and ectodermal lineage are described, as well as methods of isolating, expanding, culturing, and cryopreserving such cells.
4 Citations
33 Claims
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1. A method for isolating umbilical cord lining stem cells (ULSCs) from an umbilical cord, said method comprising
a) obtaining the lining of an umbilical cord, wherein said lining is substantially free of blood, venous tissue, and arterial tissue; - and
b) culturing explants of said lining on a fibronectin-coated solid substrate in the presence of a low glucose growth medium for a period of time sufficient for said ULSCs to adhere to said fibronectin-coated solid substrate, said growth medium comprising 15% fetal bovine serum, a stabilized dipeptide of L-alanyl-L-glutamine, antibiotic, and a growth factor selected from the group consisting of basic fibroblast growth factor (bFGF), leukemia inhibitory factor (LIF), and epidermal growth factor (EGF). - View Dependent Claims (2, 3, 4, 5, 6, 7, 8)
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9. A composition for culturing ULSCs, said composition comprising:
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a) a low glucose growth medium; b) 10% to 20% serum; c) 0.7 to 1.5% of a stabilized dipeptide of L-alanyl-L-glutamine; d) 1 to 100 ng/mL of a growth factor selected from the group consisting of basic fibroblast growth factor (bFGF), leukemia inhibitory factor (LIF), and epidermal growth factor (EGF); and e) 1 to 3% of an antibiotic. - View Dependent Claims (10, 11, 12, 13)
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14. A composition comprising a purified population of ULSCs and a culture medium, wherein said culture medium comprises a low glucose growth medium;
- 10% to 20% serum;
0.7 to 1.5% of a stabilized dipeptide of L-alanyl-L-glutamine;
1 to 100 ng/mL of a growth factor selected from the group consisting of bFGF, LIF, and EGF; and
1 to 3% of an antibiotic, wherein said ULSCs are positive for CD105, CD106, CD90, CD73, SSEA-4, and STRO-1, and negative for CD45, CD34, CD19, and HLA-DR. - View Dependent Claims (15, 16, 17)
- 10% to 20% serum;
- 18. A purified population of umbilical cord lining stem cells, wherein said cells are positive for CD105, CD106, CD90, CD73, SSEA-4, and STRO-1, negative for CD45, CD34, CD19, and HLA-DR, express OCT4 and Nanog, and do not express Sox2.
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31. A method for culturing a population of ULSCs, said method comprising obtaining a population of ULSCs from human umbilical cord, wherein said ULSCs are positive for CD105, CD106, CD90, CD73, SSEA-4, and STRO-1, negative for CD45, CD34, CD19, and HLA-DR, express OCT4 and Nanog, and do not express Sox2;
- and culturing said cells in the presence of a low glucose growth medium containing 10% to 20% serum;
0.7 to 1.5% of a stabilized dipeptide of L-alanyl-L-glutamine;
1 to 100 ng/mL of a growth factor selected from the group consisting of bFGF, LIF, and EGF; and
1 to 3% of an antibiotic. - View Dependent Claims (32, 33)
- and culturing said cells in the presence of a low glucose growth medium containing 10% to 20% serum;
Specification