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 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.
Jasmund
I
I
KTE, University Lübeck, Germany.
Schwientek
S
S
Acikgöz
A
A
Langsch
A
A
Machens
H G
HG
Bader
A
A
D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D002470:Cell Survival; D002478:Cells, Cultured; D003470:Culture Media / Q000737:chemistry* / Q000378:metabolism*; D005109:Extracellular Matrix / Q000737:chemistry / Q000378:metabolism*; D022781:Hepatocytes / Q000166:cytology* / Q000502:physiology*; D006801:Humans; D013045:Species Specificity; D013552:Swine; D023822:Tissue Engineering / Q000379:methods*
D003470:Culture Media