Conditionally adherent growth of serum-independent CHO cells for automated drug screening and biopharmaceutical production
✍ Scribed by Christine R. Gandor; Michael O. Zang-Gandor; Peter J. Flor; Marcello Marangi; Fred A.M. Asselbergs
- Book ID
- 101243068
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 485 KB
- Volume
- 65
- Category
- Article
- ISSN
- 0006-3592
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✦ Synopsis
SSF3 is a CHO cell line adapted for growth in protein-free medium. It grows in suspension unless serum-derived attachment factors such as vitronectin are added to the medium. Serum-independent cell lines, which adhere to the substrate after induction with dexamethasone or constitutively, were created by transfection with a human vitronectin gene under control of the mouse mammary tumor-virus promoter. Substrate attachment and SSF3VN-cell spreading could be prevented with an RGD peptide (arginine-glycine-aspartic acid) confirming that attachment is mediated by an intregrin receptor. Hormone-inducible attachment could be blocked by glucocorticoid antagonist promegestone. All steps in the isolation of stable transfected SSF3VN cell lines could be done in a chemically defined medium avoiding the risk of introduction of serum-derived infectious agents. SSF3VN cells could be grown in protein-free medium in solid-phase large-scale bioreactors. Application in microplates as used in high-throughput screening was demonstrated in an assay of Ca 2+ release from internal stores induced by agonist-binding to recombinant human metabotropic glutamate receptor hmGluR1b.