Biotechnology Letters

, Volume 34, Issue 8, pp 1453–1458

Novel cholesterol feeding strategy enables a high-density cultivation of cholesterol-dependent NS0 cells in linear low-density polyethylene-based disposable bioreactors

  • Yiwen Tao
  • Helena Yusuf-Makagiansar
  • Jennifer Shih
  • Thomas Ryll
  • Marty Sinacore
Original Research Paper

DOI: 10.1007/s10529-012-0915-6

Cite this article as:
Tao, Y., Yusuf-Makagiansar, H., Shih, J. et al. Biotechnol Lett (2012) 34: 1453. doi:10.1007/s10529-012-0915-6

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 × 106 viable cells/ml at ≥90 % cell viability were achieved. Vials of high density cell banks were created by filling 90–100 × 106 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.

Keywords

Cell banksCholesterolDisposable bioreactorCyclodextrinNS0 cellsPerfusion cell culture

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Yiwen Tao
    • 1
  • Helena Yusuf-Makagiansar
    • 1
  • Jennifer Shih
    • 1
  • Thomas Ryll
    • 1
  • Marty Sinacore
    • 1
  1. 1.Cell Culture Development, Biogen Idec IncCambridgeUSA