Adaptation of mammalian cells to growth in serum-free media.
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.
Sinacore
M S
MS
Genetic Institute, Andover, MA 01810, USA.
Drapeau
D
D
Adamson
S R
SR
D000222:Adaptation, Physiological; D000818:Animals; D019149:Bioreactors; D016466:CHO Cells; D002452:Cell Count; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002455:Cell Division; D002478:Cells, Cultured / Q000166:cytology* / Q000502:physiology; D006224:Cricetinae; D016895:Culture Media, Serum-Free; D011994:Recombinant Proteins / Q000096:biosynthesis*
D016895:Culture Media, Serum-Free; D011994:Recombinant Proteins