Novel cholesterol feeding strategy enables a high-density cultivation of cholesterol-dependent NS0 cells in linear low-density polyethylene-based disposable bioreactors |
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Authors: | Yiwen Tao Helena Yusuf-Makagiansar Jennifer Shih Thomas Ryll Marty Sinacore |
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Affiliation: | Cell Culture Development, Biogen Idec Inc, Cambridge, MA 02142, USA. |
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Abstract: | We have developed a perfusion-based high cell density (HD) cell banking and inoculum expansion procedure for a cholesterol-dependent NS0 myeloma cell line using linear low-density polyethylene-based disposable bioreactors. Challenges associated with cholesterol-polymer interactions, which suppress cholesterol-dependent NS0 myeloma cell growth, were overcome using a novel cholesterol feeding protocol that included a combination of two cholesterol formulations: an ethanol-based formulation and an aqueous formulation. Using a cholesterol feed optimized for HD cell culture in a disposable bioreactor perfusion system, cell densities of >25 × 10(6) viable cells/ml at ≥ 90 % cell viability were achieved. Vials of high density cell banks were created by filling 90-100 × 10(6) viable cells/ml in 5 ml cryotube vials. Implementation of the HD cell banks enabled a significant reduction in the number of step operations in the inoculum expansion phase in a large-scale manufacturing setting. |
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