title: Novel cholesterol feeding strategy enables a high-density cultivation of cholesterol-dependent NS0 cells in linear low-density polyethylene-based disposable bioreactors.
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.
authors.affiliation: Cell Culture Development, Biogen Idec Inc, Cambridge, MA 02142, USA.
authors.forename: Yiwen
authors.initials: Y
authors.lastname: Tao
authors.forename: Helena
authors.initials: H
authors.lastname: Yusuf-Makagiansar
authors.forename: Jennifer
authors.initials: J
authors.lastname: Shih
authors.forename: Thomas
authors.initials: T
authors.lastname: Ryll
authors.forename: Marty
authors.initials: M
authors.lastname: Sinacore
mesh_terms: D000818:Animals; D019149:Bioreactors; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D045744:Cell Line, Tumor; D002470:Cell Survival; D002784:Cholesterol / Q000737:chemistry / Q000378:metabolism*; D003470:Culture Media / Q000737:chemistry / Q000378:metabolism; D000431:Ethanol / Q000737:chemistry / Q000378:metabolism; D007700:Kinetics; D051379:Mice; D020959:Polyethylene
chemical_list: D003470:Culture Media; D000431:Ethanol; D020959:Polyethylene; D002784:Cholesterol