Molecular Biotechnology

, Volume 15, Issue 3, pp 249–257

Adaptation of mammalian cells to growth in serum-free media

Authors

  • Martin S. Sinacore
    • Genetic Institute
  • Denis Drapeau
  • S. R. Adamson
Protocol

DOI: 10.1385/MB:15:3:249

Cite this article as:
Sinacore, M.S., Drapeau, D. & Adamson, S.R. Mol Biotechnol (2000) 15: 249. doi:10.1385/MB:15:3:249

Abstract

A three-step protocol is described for adapting an anchorage-dependent, serum-dependent recombinant mammalian cell lineage to high density serum-free suspension culture. The objective is a cell lineage that is well-suited for the manufacture of a recombinant protein. The first step of the protocol generates an anchorage-independent cell lineage by culturing trypsin-treated cells in spinner flasks using serum-containing medium. The second step adapts the lineage to serum-free medium through a series of serum reduction steps in the presence of defined growth-promoting additives. The third step adapts the lineage to high-cell-density conditions by culturing the cells in a bioreactor in a manner that allows development of tolerance to growth-inhibiting substances released by the cells. Examples are presented for the use of this protocol for recombinant CHO cells.

Index Entries

Mammalian cell cultureserum-free mediumsuspension cultureadaptationbiopharmaceuticalbioreactor

Copyright information

© Humana Press Inc 2000