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Application of collagen coated silicone scaffolds for the three‐dimensional cell culture of primary rat hepatocytes
Author(s) -
Stock Peggy,
Winkelmann Claudia,
Thonig Antje,
Böttcher Gunther,
Wenske Günther,
Christ Bruno
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.274.2
Subject(s) - silicone , scaffold , hepatocyte , cell culture , chemistry , primary culture , 3d cell culture , urea , cytotoxicity , cell growth , microbiology and biotechnology , biomedical engineering , cell , biology , biochemistry , medicine , in vitro , organic chemistry , genetics
Primary hepatocytes present an appropriate validation system for toxicological studies of novel agents in pharmaceutical‐ and cosmetics industries. Until today, three dimensional culture systems maintaining the functionality of hepatocytes are missing. The aim was to analyse the applicability of 3D collagen‐coated silicone scaffolds versus 2D culture for culture of rat hepatocytes. Primary rat hepatocytes were cultivated for 72h on multilayer lattice structures (A=23.7cm 2 ) and compared with 2D culture on conventional dishes (A=8.9cm 2 ). Cell counts, the growth pattern on silicone and the urea production rate were determined. Based on identical initial cell counts the growth rate was about 42%±5% higher in 3D culture as compared to 2D. Under serum‐free conditions cells attached to the scaffold, formed clusters and organ‐like structures. The 3D culture on the silicone scaffold displayed no significant cytotoxicity after 72h: the percentages of dead cells were 7%±3% vs 9%±3%. The urea production rate [μg/10 5 cells] was significantly higher in the 3D vs 2D: 24h 9.1±2.7 vs 4.7±0.2; 48h 5.7±1.5 vs 3.7±0.05 72h 3.5±0.5 vs 2.1±0.1. The results show the appropriateness of silicone scaffolds for the 3D culture of primary rat hepatocytes. Thus, the collagen‐coated silicone scaffolds represent an important tool for the development of 3D hepatocyte cultures sustaining the physiological functions.