Chemically defined cell culture media and system and methods for use, particularly for culturing epithelial cells
AssigneeOrganogenesis, Inc.
InventorsNancy Louise Parenteau, Eric William Johnson, Susan Frances Meunier, John Gregory Maresh
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Abstract
The present invention provides systems, methods and chemically defined media for the cultivation of cells, particularly epithelial cells. Cells may be cultured with a varied calcium concentration. Furthermore, a calcium concentation in excess of 1.00 mM may be used in the practice of the present invention without loss of a proliferative cell population and maintianing high colony forming efficiencies. Population doubling times range from about 16 to about 33 hours. Cells may serially cultivated to achieve from about 20 to about 50 population doublings.
Claims27
A chemically defined cell culture medium comprising, insulin or an insulin-like growth factor;
- transferrin or ferrous ion;
- triiodothyronine or thyroxin;
- at least one ethanolamine or o-phosphoryl-ethanolamine;
- calcium;
- and a nutrient source in the absence of undefined components, wherein:
- (a) Insulin is present at from about 0.5 to about 50 .mu.g/ml or an insulin-like growth factor is present at from about 10.sup.-10 to about 10.sup.-8 M;
- (b) Transferrin is present at from about 0.05 to about 50 .mu.g/ml or ferrous ion is present at from about 5.times.10.sup.-8 to about 5.times.10.sup.-5 M;
- (c) Triiodothyronine is present at from about 2 to about 200 .rho.M;
- (d) At least one of o-phosphoryl-ethanolamine and ethanolamine is present at from about 10.sup.-6 to about 10.sup.-2 M;
- (e) Calcium is present at from about 0.005 to about 2.0 mM;
- and
- (f) Nutrient source is selected from at least one of DMEM, IDMEM, MEM, M199, RPMI 1640, Ham's F12, Ham's F10, NCTC 109, and NCTC 135.
Dependencies are inferred from the claim text.