Ultra-high density cell banking methods
AssigneeGenzyme Corporation
InventorsXiaoxia Jin, Claudia Buser
Life of this patent
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Abstract
Provided are methods for the creation of ultra-high density cryopreserved cell banks. In certain embodiments, these methods employ altered perfusion culture techniques that allow for production of ultra-high density cell cultures that can be cryopreserved at unexpectedly high cell densities without the need for any cell concentration steps, while retaining excellent cell viability and quality.
Claims21
A method for producing an ultra-high density frozen mammalian cell bank directly from a population of cultured cells, the method comprising:
- a) culturing the mammalian cells in a perfusion bioreactor to obtain the ultra-high density cell population with a concentration of at least about 1.0×108 cells/mL, wherein said perfusion bioreactor is coupled to a cell retention system;
- b) harvesting the ultra-high density population directly without performing a concentration step after culturing the mammalian cells in the perfusion bioreactor that is coupled to the cell retention system;
- and
- c) cryopreserving the harvested ultra-high density cell population by adding a cryoprotectant to produce an ultra-high density frozen cell bank,
- wherein the ultra-high density frozen cell bank has a concentration of at least about 1.0×108 cells/mL and at least 95% post-thaw viability.
A method for producing a culture of mammalian cells from an ultra-high density frozen cell bank, the method comprising thawing and culturing the ultra-high density frozen cell bank with a concentration of at least about 1.0×108 cells/mL in a perfusion bioreactor to obtain the culture of mammalian cells,
- wherein the ultra-high density frozen cell bank was produced by harvesting a cell culture directly from a bioreactor, wherein said bioreactor is coupled to a cell retention system, without performing any concentration steps after culturing the mammalian cells in the perfusion bioreactor and cryopreserving the harvested cell culture after adding a cryoprotectant, and wherein the cultured mammalian cells have at least 95% post-thaw viability.
A method for producing an ultra-high density frozen cell bank, the method comprising:
- a) harvesting a culture of mammalian cells directly from a bioreactor, wherein the bioreactor is coupled to a non-centrifugal cell retention device, and wherein the harvesting is conducted without performing any concentration step after culturing the mammalian cells in the bioreactor that is coupled to the non-centrifugal cell retention device;
- and
- b) cryopreserving the harvested culture by adding a cryoprotectant to produce the ultra-high density frozen cell bank,
- wherein the ultra-high density frozen cell bank has a concentration of at least about 1.0×108 cells/mL and at least 95% post-thaw viability in a perfusion culture.
Dependencies are inferred from the claim text.