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US 2014/0273206 A1Published application
High density cell banking methods
AssigneeGenzyme Corporation
InventorsXiaoxia JIN, Haodi DONG, Claudia BUSER
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Abstract
The current disclosure provides a method for the creation of a high-density cryopreserved cell bank using perfusion culture techniques and non-centrifugal concentration of cells. Methods of production using this high-density cryopreserved cell bank are also provided.
Claims36
A non-centrifugal method for producing a high-density frozen cell bank, the method comprising:
- a) culturing cells in a perfusion bioreactor, wherein said bioreactor is coupled to a cell retention system;
- b) non-centrifugally concentrating said cells to produce a concentrated cell population;
- c) cryopreserving the concentrated cell population to produce a high-density frozen cell bank.
A non-centrifugal method for producing a high-density frozen cell bank, the method comprising:
- a) culturing cells in a perfusion bioreactor coupled to an alternating tangential flow filtration system, wherein the bioreactor comprises a flexible bag bioreactor, and wherein the filter has a filter surface area of at least 0.3 m2 and a filter with a MWCO size of at least 50 kDa;
- b) non-centrifugally concentrating the cells using the alternating tangential flow filtration system to produce a concentrated cell population having a density of about 1×10̂8 cells/mL;
- c) cryopreserving the concentrated cell population to produce a high-density frozen cell bank, wherein the cryopreserving comprises adding DMSO to the concentrated cell population to a final concentration of about 5% to aboutl0%, vol/vol;
- and wherein the high-density frozen cell bank has a cell density of about 10̂8 cells/mL.
1 more element
Dependencies are inferred from the claim text.