Serum-free cell growth medium
AssigneeThe United States of America as represented by the Department of Health and Human Services
InventorsFrank P. Luyten, Ludwig Erlacher
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Abstract
A chemically defined-serum free growth medium for the in vitro and ex vivo of cells and cell lines. The medium consists of about a one to one ratio (v/v) of two basal growth media containing α-ketoglutarate, insulin, transferrin, selenium, bovine serum albumin, linoleic acid, ceruloplasmin, cholesterol, phosphatidyl-ethanolamine, α-tocopherol acid succinate, reduced glutathione, taurine, triiodothyronine, hydrocortisone, parathyroid hormone, L-ascorbic acid 2-sulfate, β-glycerophosphate, PDGF, EGF and FGF. Chondrocytes, when cultured in this medium in the presence of a cartilage derived morphogenetic protein or bone morphogenetic protein, retain their cartilaginous phenotype.
Claims20
A method of maintaining a cartilaginous phenotype in chondrocytes in vitro comprising contacting a culture comprising chondrocytes, which have not lost and subsequently re-expressed a cartilaginous phenotype, with a serum-free cell growth medium comprising about a 1:1 ratio (v/v) of two basal cell culture media, said medium containing effective cell growth-promoting concentrations of α-ketoglutarate, insulin, transferrin, selenium, bovine serum albumin, linoleic acid, ceruloplasmin, cholesterol, phosphatidylethanolamine, α-tocopherol acid succinate, reduced glutathione, taurine, triiodothyronine, hydrocortisone, parathyroid hormone, L-ascorbic acid 2-sulfate, β-glycerophosphate, platelet-derived growth factor (PDGF), epidermal growth factor (EGF) and basic fibroblast growth factor (bEGF) and at least one morphogenetic protein selected from the group consisting of cartilage-derived morphogenetic proteins and bone morphogenetic proteins, whereby a cartilaginous phenotype is maintained in said chondrocytes in vitro.
A method of repairing a joint surface defect in a mammal in need thereof comprising the steps of:
- removing normal cartilage in the vicinity of said surface defect;
- isolating chondrocytes from said cartilage;
- contacting a culture comprising said chondrocytes, which have not lost and subsequently re-expressed a cartilaginous phenotype, with a serum-free cell growth medium comprising about a 1:1 ratio (v/v) of two basal cell culture media, said medium containing effective cell growth-promoting concentrations of α-ketoglutarate, insulin, transferrin, selenium, bovine serum albumin, linoleic acid, ceruloplasmin, cholesterol, phosphatidylethanolamine, α-tocopherol acid succinate, reduced glutathione, taurine, triiodothyronine, hydrocortisone, parathyroid hormone, L-ascorbic acid 2-sulfate, β-glycerophosphate, platelet-derived growth factor (PDGF), epidermal growth factor (EGF) and basic fibroblast growth factor (bEGF) and at least one morphogenetic protein selected from the group consisting of cartilage-derived morphogenetic proteins and bone morphogenetic proteins, whereby a cartilaginous phenotype is maintained in said chondrocytes;
- and
- implanting said chondrocytes into said surface defect.
Dependencies are inferred from the claim text.