Abstract
A rapid and effective semi-automated screening method has been developed for the development of growth media for mammalian cell culture. The method has proven to be a powerful tool for preliminary evaluation and comparison of new media formulations, but has some inherent disadvantages, which are important to recognise in order to interpret the results.
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Abbreviations
- AUC:
-
Area under curve
- DMEM:
-
Dulbecco’s modified eagles medium
- DMF:
-
N,N-Dimethylformamide
- ELISA:
-
Enzyme linke immunosorbent assay
- FCS:
-
Fetal calf serum
- HEPES:
-
Hydroxy-ethyl-piperazine-ethane-sulphonic acid
- MTT:
-
3-(4,5-Dimethylthiazol-2-yl)2,5-diphenyl-tetrazolium bromide
- PEG:
-
Polyethylen glycol
- SDS:
-
Sodium dodecylsulphate
References
Alley Cet al. (1988) Feasibility of drug screening with panels of human tumor cell lines using a microculture tetrazolium assay. Cancer Research 48: 589–601.
Al-Rubeai M and Spier R (1989) Use of the MTT assay for the study of hybridoma cells in homogeneous and heterogeneous cultures. In: Spier RE, Griffiths JB, Stephenne J and Crooy PJ (eds.) Advances in Animal Cell Biology and Technology for Bioprocesses. (pp. 143–155) Butterworth, London.
Altman FP (1977) Tetrazolium salts and formazans. In: Progress in histochemistry and cytochemistry vol. 9 (pp. 1–56) Gustav Fishers Verlag, Stuttgart.
Andresen LO, Koch C, Sørensen BB and Emborg C (1989) Production of monoclonal antibodies against the enzyme nuclease fromSerratia marcecens and evaluation of the individual antibodies for their use in EIA and in affinity chromatography. Biotechnol. Techniques 3: 407–410.
Box GEP, Hunter WG and Hunter JS (1978) Statistics for experimenters. John Wiley & Sons, New York.
Denizot F and Lang R (1986) Rapid colorimetric assay for cell growth and survival. J. Immunol. Methods 89: 271–277.
Franěk F and Dolníková J (1991) Hybridoma growth and monoclonal antibody production in iron-rich protein-free medium: Effect of nutrient concentration. Cytochnology 7: 33–38.
Gerlier D and Thomasset N (1986) Use of MTT colorimetric assay to measure cell activation. J. Immunol. Methods 94: 57–63.
Hansen MB, Nielsen SE and Berg K (1989) Re-examination and further development of a precise and rapid dye method for measuring cell growth/cell kill. J. Immunol. Methods 119: 203–210.
Hicks CR (1982) Fundamental concepts in the design of experiments. 3rd Ed. Holt Saunders, Tokyo
Kierulff JV (1991) Development of serum-free media for mammalian cell culture. Ph.D. dissertation, Department of Biotechnology, Technical University of Denmark.
Luan LY, Mutharasan R and Magee WE (1987) Effect of various glucose/glutamine ratios on hybridoma growth, viability and monoclonal antibody formation. Biotechnol. Lett. 9: 535–538.
Marquis CPet al. (1989) Agitation and aeration effects in suspension mammalian cell cultures. Cytotechnology 2: 163–170.
Mignot G, Ganne V, Faure T, Pavirani A and Van de Pol H (1989) The use of a statistical approach for the optimization of culture conditions of genetically engineered cell lines. In: Spier RE, Griffiths JB, Stephenne J and Crooy PJ (eds.) Advances in animal cell biology and technology for bioprocesses. (pp. 52–58) Butterworth, London.
Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods 65: 55–63.
Twentyman PR and Luscombe M (1987) A study of some variables in a tetrazolium dye (MTT) based assay for cell growth and chemosensitivity. Br. J. Cancer 56: 279–285.
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Rexen, P., Valentin Kierulff, J. & Emborg, C. An easy-to-handle semi-automated method for media development using a colorimetric viability assay and fractional factorial designs. Cytotechnology 8, 195–205 (1992). https://doi.org/10.1007/BF02522036
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DOI: https://doi.org/10.1007/BF02522036