The influence of medium composition and matrix on long-term cultivation of primary porcine and human hepatocytes

The differentiated hepatocyte phenotype remains difficult to maintain in culture. The duration over which phenotypically stable hepatocytes can be cultured ranges from a couple of days to a few weeks. Shortcomings in medium formulation may be a factor in this lack of success. We have investigated ef...

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Bibliographic Details
Published in:Biomolecular engineering 2007-02, Vol.24 (1), p.59-69
Main Authors: Jasmund, I., Schwientek, S., Acikgöz, A., Langsch, A., Machens, H.G., Bader, A.
Format: Article
Language:English
Subjects:
Animals
Cell Culture Techniques - methods
Cell Proliferation
Cell Survival
Cells, Cultured
Culture Media - chemistry
Culture Media - metabolism
Extracellular Matrix - chemistry
Extracellular Matrix - metabolism
Fibronectin
Hepatocytes - cytology
Hepatocytes - physiology
Human
Humans
Matrix
Medium composition
Porcine
Primary hepatocytes
Serum
Species Specificity
Swine
Tissue Engineering - methods
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Summary:The differentiated hepatocyte phenotype remains difficult to maintain in culture. The duration over which phenotypically stable hepatocytes can be cultured ranges from a couple of days to a few weeks. Shortcomings in medium formulation may be a factor in this lack of success. We have investigated effects of medium formulation on primary porcine and human hepatocyte cultures. We tested seven culture medium compositions (DMEM, ExCell 400, HepatoZYME-SFM, L-15 Leibovitz, SF-3, Waymouth, and Williams’ E) and the effects of serum, fibronectin and biomatrix in a sandwich culture configuration. Albumin, urea, cholesterol, GOT, GPT, LDH and triglyceride concentrations were measured over 14 days. For both human and porcine cultures, the best results were obtained with SF-3 medium. Cells cultivated with Williams’ E medium and FCS had good morphology and synthetic function during the first days of culture. However, continued addition of serum, was associated with a subsequent loss of differentiated phenotype. Addition of fibronectin was associated with improved function in cultures maintained in SF-3 medium whilst biomatrix had no effect. In contrast, addition of fibronectin did not influence cultures maintained in Williams’ E medium, but cultures with biomatrix were associated with improved function at longer time points.
ISSN:1389-0344
1878-559X
DOI:10.1016/j.bioeng.2006.05.018