Conditionally adherent growth of serum-independent CHO cells for automated drug screening and biopharmaceutical production

Autor: Michael O. Zang-Gandor, Christine Gandor, Peter J. Flor, Marcello Marangi, Fred A.M. Asselbergs
Rok vydání: 1999
Předmět:
Zdroj: Biotechnology and Bioengineering. 65:523-528
ISSN: 1097-0290
0006-3592
DOI: 10.1002/(sici)1097-0290(19991205)65:5<523::aid-bit4>3.0.co;2-6
Popis: 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.
Databáze: OpenAIRE
Externí odkaz: