Adaptation of mammalian cells to growth in serum-free media

被引：95

作者：

Sinacore, MS ^{[1
]}

Drapeau, D ^{[1
]}

Adamson, SR ^{[1
]}

机构：

[1] Genet Inst Inc, Andover, MA 01810 USA

来源：

MOLECULAR BIOTECHNOLOGY | 2000年 / 15卷 / 03期

关键词：

mammalian cell culture; serum-free medium; suspension culture; adaptation; biopharmaceutical; bioreactor;

D O I：

10.1385/MB:15:3:249

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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.

引用

页码：249 / 257

页数：9

共 37 条

[1]

ADAMSON SR, 1996, ADAPTATION MAMMALIAN

[2]

ASSOIAN RK, 1977, J CELL BIOL, V136, P1

[3] METHODS FOR GROWTH OF CULTURED-CELLS IN SERUM-FREE MEDIUM [J].