Development of a serum-free medium for the production of erythropoietin by suspension culture of recombinant Chinese hamster ovary cells using a statistical design

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In order to develop a serum-free (SF) medium for the production of erythropoietin (EPO) by suspension culture of recombinant Chinese hamster ovary (rCHO) cells, a statistical optimization approach based on a Plackett-Burman design was adopted. A basal medium was prepared by supplementing Iscove's modified Dulbecco's medium (IMDM) with Fe(NO3)(3)(.)9H(2)O, CuCl2 and ZnSO(4)(.)7H(2)O which are generally contained in SF medium formulations. Insulin, transferrin and ethanolamine were also supplemented to the basal medium to determine their optimal concentrations. From this statistical analysis, glutamate, serine, methionine, phosphatidylcholine, hydrocortisone and pluronic F68 were identified as positive determinants for cell growth. The SF medium was formulated by supplementing the basal medium with components showing positive effects on cell growth in suspension culture. An EPO titer in this optimized SF medium was 79% of that in IMDM supplemented with 5% dialyzed fetal bovine serum (dFBS). Furthermore. the in vitro and in vivo biological activities of EPO produced in the SF medium were comparable to those produced in the serum-supplemented medium. Taken together, the results obtained hen show that a Plackett-Burman design facilitates the development of SF media for the production of EPO by suspension culture of rCHO cells. (C) 1999 Elsevier Science B.V. All rights reserved.
Publisher
ELSEVIER SCIENCE BV
Issue Date
1999-04
Language
English
Article Type
Article
Keywords

INTERFERON-GAMMA PRODUCTION; ANIMAL-CELLS; GROWTH; OPTIMIZATION; EXPRESSION

Citation

JOURNAL OF BIOTECHNOLOGY, v.69, no.2-3, pp.85 - 93

ISSN
0168-1656
URI
http://hdl.handle.net/10203/72067
Appears in Collection
BS-Journal Papers(저널논문)
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