Culture of HepG2 liver cells on three dimensional polystyrene scaffolds enhances cell structure and function during toxicological challenge.
Cultured cells are dramatically affected by the micro-environment in which they are grown. In this study, we have investigated whether HepG2 liver cells grown in three dimensional (3-D) cultures cope more effectively with the known cytotoxic agent, methotrexate, than their counterparts grown on traditional two dimensional (2-D) flat plastic surfaces. To enable 3-D growth of HepG2 cells in vitro, we cultured cells on 3-D porous polystyrene scaffolds previously developed in our laboratories. HepG2 cells grown in 3-D displayed excellent morphological characteristics and formed numerous bile canaliculi that were seldom seen in cultures grown on 2-D surfaces. The function of liver cells grown on 3-D supports was significantly enhanced compared to activity of cells grown on 2-D standard plasticware. Unlike their 2-D counterparts, 3-D cultures were less susceptible to lower concentrations of methotrexate. Cells grown in 3-D maintained their structural integrity, possessed greater viability, were less susceptible to cell death at higher levels of the cytotoxin compared to 2-D cultures, and appeared to respond to the drug in a manner more comparable to its known activity in vivo. Our results suggest that hepatotoxicity testing using 3-D cultures might be more likely to reflect true physiological responses to cytotoxic compounds than existing models that rely on 2-D culture systems. This technology has potential applications for toxicity testing and drug screening.
Bokhari
Maria
M
School of Biological and Biomedical Science, Durham University, South Road, Durham DH1 3LE, UK.
Carnachan
Ross J
RJ
Cameron
Neil R
NR
Przyborski
Stefan A
SA
D002448:Cell Adhesion; D002450:Cell Communication; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor / Q000166:cytology* / Q000648:ultrastructure; D006801:Humans; D021621:Imaging, Three-Dimensional; D008099:Liver / Q000166:cytology* / Q000648:ultrastructure; D008727:Methotrexate / Q000633:toxicity; D008854:Microscopy, Electron; D011137:Polystyrenes; D018675:Toxicity Tests / Q000379:methods
D011137:Polystyrenes; D008727:Methotrexate
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