Abstract
Stepwise screening of media supplements using factorial design and analysis was employed in the development of serum-free medium for a recombinant Chinese hamster ovary cell line. The effects (growth and target protein production) of different combinations were measured at two time points to ensure adequate response. The results were analysed by a computer program specialized in factorial analysis. The formulation deduced from the previous experiment was used as the new basal medium for the next screening. Certain significant nutrients were studied again in a more ‘advanced’ formulation in order to analyse the potential synergistic effects with new media components. Compared to cells grown in serum-containing medium, cells adapted to the final formulation of the serum-free medium had a comparable growth rate but a four fold increase in the active protein production.
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Abbreviations
- ANOVA:
-
Analysis of variance
- BSA:
-
bovine serum albumin
- CHO:
-
Chinese hamster ovary
- FBS:
-
fetal bovine serum
- MTT:
-
3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide
- PBS:
-
phosphate buffered saline
- SFM:
-
serum-free medium
References
Barnes D and Sato G (1980) Methods for growth of cultured cells in serum-free medium. Analytical Biochemistry 102: 255–270.
Kierulff J, Persson B, Rexen P, Morcel C and Emborg C (1993) Development of serum-free media for mammalian cell culture. ESACT 12th Meeting pp. 133–135.
Montgomery D (1991) Design and analysis of experiments. 3rd Ed. John Wiley & Sons, New York.
Wallace L, McKeehan W, Gregory H and Ham R (1976) Selenium is an essential trace nutrient for growth of WI-38 diploid human fibroblasts. Proc. Natl. Acad. Sci. USA 73(6): 2023–2027.
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Lao, MS., Schalla, C. Development of a serum-free medium using computer-assisted factorial design and analysis. Cytotechnology 22, 25–31 (1996). https://doi.org/10.1007/BF00353921
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DOI: https://doi.org/10.1007/BF00353921