Cytotechnology

, Volume 57, Issue 3, pp 251–261

Evaluation and optimization of hepatocyte culture media factors by design of experiments (DoE) methodology

Authors

  • Jia Dong
    • Division of Biotechnology/IFMLinköping University
    • Division of Biotechnology/IFMLinköping University
  • Marc Lübberstedt
    • Department of SurgeryCharite Universitätsmedizin Berlin
  • Thomas Urbaniak
    • Department of SurgeryCharite Universitätsmedizin Berlin
  • Andreas K. N. Nüssler
    • Department of TraumatologyTU Munich, MRI
  • Daniel Knobeloch
    • Department of SurgeryCharite Universitätsmedizin Berlin
  • Jörg C. Gerlach
    • Department of SurgeryCharite Universitätsmedizin Berlin
    • Berlin Brandenburg Center for Regenerative Therapies (BCRT)Charite Universitätsmedizin Berlin
    • Departments of Surgery and Bioengineering, McGowan Institute for Regenerative MedicineUniversity of Pittsburgh
  • Katrin Zeilinger
    • Department of SurgeryCharite Universitätsmedizin Berlin
    • Berlin Brandenburg Center for Regenerative Therapies (BCRT)Charite Universitätsmedizin Berlin
Original Research

DOI: 10.1007/s10616-008-9168-6

Cite this article as:
Dong, J., Mandenius, C., Lübberstedt, M. et al. Cytotechnology (2008) 57: 251. doi:10.1007/s10616-008-9168-6

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.

Keywords

Design of experimentsBioreactorC3APrimary human hepatocytesGrowth factorsCulture medium optimization

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

Copyright information

© Springer Science+Business Media B.V. 2008