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Cytotechnology

, Volume 37, Issue 3, pp 189–198 | Cite as

Development and improvement of a serum-free suspension process for the production of recombinant adenoviral vectors using HEK293 cells

  • Yung-Shyeng Tsao
  • Russell Condon
  • Eugene Schaefer
  • Peggy Lio
  • Zhong Liu
Article

Abstract

Human Embryonic Kidney 293 (HEK293) cells were adapted into a serum-free suspension medium through steps of gradual serum weaning for the production of adenoviral (AdV) gene therapy vectors. The presence of sodium heparin in the medium formulation reduced cell clumping dramatically in suspension culture. The adapted cells were ready to grow either in serum-containing medium as an attached culture or in serum-free medium in suspension culture. A scalable production process was developed in shake flasks and was then evaluated in stirred tank bioreactors. This process includes a growth phase in batch-mode followed by a production phase involving medium perfusion and supplementation. Fortification with calcium chloride post viral inoculation resulted in an increase in virus production by at least one fold. Addition of stimulating agents such as sodium butyrate, N-acetyl-L-cysteine (NAC), dimethyl sulfoxide(DMSO), or ethyl alcohol post infection was shown to further improve virus production in a dose-dependent manner. The serum-free suspension process described here should be suitable for the manufacturing of other E1-deleted AdV vectors and could potentially be used for the production of recombinant proteins by HEK293 cells.

Adenovirus Dimethyl sulfoxide (DMSO) Ethyl alcohol Gene therapy Human Embryonic Kidney 293 (HEK293) cells N-acetyl-L-cysteine Recombinant adenoviral vector Serum-free suspension culture Sodium butyrate 

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Copyright information

© Kluwer Academic Publishers 2001

Authors and Affiliations

  • Yung-Shyeng Tsao
    • 1
  • Russell Condon
    • 1
  • Eugene Schaefer
    • 1
  • Peggy Lio
    • 1
  • Zhong Liu
    • 1
  1. 1.Biotechnology DevelopmentSchering-Plough Research InstituteUnionUSA

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