Adaption of mammalian cell lines to high cell densities
AssigneeGenetics Institute, Inc.
InventorsS. Robert Adamson, Denis Drapeau, Yen-Tung Luan, Douglas A. Miller
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Abstract
Methods and nutrient media are disclosed useful for adapting mammalian cell lines to culture at increased cell densities.
Claims9
A method for producing an adapted mammalian cell line which grows at increased cell densities, said method comprising:
- a) initiating a passage by diluting a culture containing mammalian cells with a suitable growth medium, the dilution factor being suitable for the duration of the passage;
- b) maintaining pH, dissolved oxygen, and nutrients at non-limiting levels during the passage;
- c) repeating steps (a) and (b) at least about 5 times;
- and
- d) obtaining an adapted mammalian cell line with the ability to grow at increased cell densities.
A method for producing an adapted CHO cell line which grows at increased cell densities, said method comprising:
- a) initiating a passage approximately 1 to 5 days in duration by diluting a culture containing CHO cells at a density of at least approximately 1.times.10.sup.6 cells/ml with a suitable growth medium, the dilution factor being suitable for the duration of the passage;
- b) maintaining pH, dissolved oxygen, and nutrients in non-limiting levels during the passage;
- c) repeating steps (a) and (b) at least about 5 times;
- and
- d) obtaining an adapted CHO cell line with the ability to grow at increased cell densities.
A method for producing an adapted CHO cell line which grows at increased cell densities, said method comprising:
- a) initiating a passage approximately 3 to 4 days in duration by diluting a culture containing CHO cells at a density of at least approximately 1.times.10.sup.6 cells/ml with a suitable growth medium, the dilution factor being suitable to the duration of the passage;
- b) maintaining pH, dissolved oxygen, and nutrients at non-limiting levels during the passage;
- c) repeating steps (a) and (b) at least about 5 times;
- and
- d) obtaining an adapted CHO cell line with the ability to grow at increased cell densities.
A method for adapting mammalian cell lines to culture at increased cell densities, said method comprising continuously or periodically diluting a cell culture, containing mammalian cells, with a suitable growth medium, for between approximately 10 to 60 days, while maintaining pH, dissolved oxygen and nutrients at non-limiting levels.
A method for adapting CHO cell lines to culture at increased cell densities, said method comprising continuously or periodically diluting a culture containing CHO cells, at a density of at least approximately 1.times.10.sup.6 cells/ml with a suitable growth medium, at a dilution rate less than approximately 0.029 hr.sup.-1, while maintaining pH, dissolved oxygen and nutrients at non-limiting levels.
Dependencies are inferred from the claim text.