Serum- and protein-free media formulations for the Chinese hamster ovary cell line DUKXB11.
The production of therapeutic proteins in mammalian cell lines is of outstanding importance. The maintenance of most mammalian cell lines in culture requires the addition of serum to the culture medium. The elimination of serum from mammalian cell culture is desirable since serum is expensive and a source of contaminants, e.g. viruses, mycoplasma or prions. Here we describe the composition of serum- and protein-free media for the Chinese hamster ovary (CHO) cell line DUKXB11. The serum-free formulation supports excellent growth of CHO DUKXB11 cells at low (23cells/cm2) and high (2 x 10(4) cells/cm2) seeding densities characterized by a generation time of 10-12h, and, after addition of 0.2% pluronic F-68, the growth of a recombinant suspension cell line derived from DUKXB11. In addition, this formulation also allowed us to adapt recombinant cell lines expressing various amounts of human antithrombin ATIII (ATIII) to serum-free conditions. Secretion of ATIII was readily observed in the serum-free medium. Minor changes to the serum-free formulation resulted in a protein free formulation that supported growth of CHO DUKXB11 cells, growth of recombinant CHO cells expressing ATIII, and production of ATIII.
Institut für Biochemie, Technische Universität Darmstadt, Petersenstr. 22, D-64287 Darmstadt, Germany. meschrod@umich.edu
Martin
M
Schröder
Kathrin
K
Matischak
Peter
P
Friedl
D000818:Animals; D000990:Antithrombin III / Q000096:biosynthesis* / Q000235:genetics; D016466:CHO Cells / Q000166:cytology* / Q000378:metabolism*; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D006224:Cricetinae; D003412:Cricetulus; D016895:Culture Media, Serum-Free / Q000737:chemistry* / Q000378:metabolism*; D006801:Humans; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis*
D016895:Culture Media, Serum-Free; D011994:Recombinant Proteins; D000990:Antithrombin III