Abstract
Optimization of cell culture media based on statistical experimental design methodology is a widely used approach for improving cultivation conditions. We applied this methodology to refine the composition of an established culture medium for growth of a human hepatoma cell line, C3A. A selection of growth factors and nutrient supplements were systematically screened according to standard design of experiments (DoE) procedures. The results of the screening indicated that the medium additives hepatocyte growth factor, oncostatin M, and fibroblast growth factor 4 significantly influenced the metabolic activities of the C3A cell line. Surface response methodology revealed that the optimum levels for these factors were 30 ng/ml for hepatocyte growth factor and 35 ng/ml for oncostatin M. Additional experiments on primary human hepatocyte cultures showed high variance in metabolic activities between cells from different individuals, making determination of optimal levels of factors more difficult. Still, it was possible to conclude that hepatocyte growth factor, epidermal growth factor, and oncostatin M had decisive effects on the metabolic functions of primary human hepatocytes.
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Abbreviations
- DexM:
-
Dexamethasone
- DoE:
-
Design of experiments
- EGF:
-
Epidermal growth factor
- FGF4:
-
Fibroblast growth factor 4
- HGF:
-
Hepatocyte growth factor
- HSA:
-
Human serum albumin
- LDH:
-
Lactate dehydrogenase
- NicA:
-
Nicotinamide
- OSM:
-
Oncostatin M
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Acknowledgments
The study was performed within the EU FP6 STREP-project Vitrocellomics. The work on primary hepatocytes was partly funded by the Federal Ministry of Education and Research (BMBF, FKZ 01GG0732 and 01GG0731 to A.N.). The authors would like to thank Dr. Ursula Müller-Vieira, Pharmacelsus GmbH, Saarbrücken, Germany, for valuable contributions.
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Dong, J., Mandenius, CF., Lübberstedt, M. et al. Evaluation and optimization of hepatocyte culture media factors by design of experiments (DoE) methodology. Cytotechnology 57, 251–261 (2008). https://doi.org/10.1007/s10616-008-9168-6
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DOI: https://doi.org/10.1007/s10616-008-9168-6