title: Spheroid organization kinetics of H35 rat hepatoma model cell system on elastin-like polypeptide-polyethyleneimine copolymer substrates.
abstract: Though two-dimensional systems have yielded some success in deriving morphological and functional markers of hepatocyte culture, they largely fail to capture the three-dimensional organization, long-term viability, and functionality of the hepatic tissue. We have engineered a system for inducing self-assembly of model H35 rat hepatoma spheroids using a copolymer comprised of biocompatible elastin-like polypeptide (ELP) chemically conjugated to positively charged polyethyleneimine (PEI). We have achieved a conjugation ratio of 30 mol %, though our studies analyzing spheroid organization kinetics indicate conjugate ratios of 5 mol % and greater to be optimal for cell culture based on least variability in spheroid sizes and minimum incidence of overgrown aggregates. Furthermore, our ELP-PEI system indicated the potential for influencing ultimate spheroid dimensions, with spheroid size inversely related to polyelectrolyte conjugation. Overall, this study provides a good starting point to investigate functional correlations between spheroid size and functional markers and their future use as an in vitro diagnostic or tissue engineering tool.
authors.affiliation: Department of Biomedical Materials Science, School of Dentistry, University of Mississippi Medical Center, Jackson, Mississippi.
authors.forename: Paul A
authors.initials: PA
authors.lastname: Turner
authors.forename: C Andrew
authors.initials: CA
authors.lastname: Weeks
authors.forename: Austin J
authors.initials: AJ
authors.lastname: McMurphy
authors.forename: Amol V
authors.initials: AV
authors.lastname: Janorkar
mesh_terms: D000818:Animals; D001672:Biocompatible Materials / Q000737:chemistry; D018929:Cell Culture Techniques / Q000379:methods*; D004549:Elastin / Q000737:chemistry*; D022781:Hepatocytes / Q000166:cytology*; D007700:Kinetics; D008099:Liver / Q000166:cytology; D010455:Peptides / Q000737:chemistry*; D011094:Polyethyleneimine / Q000737:chemistry*; D051381:Rats; D018874:Spheroids, Cellular / Q000166:cytology*; D054457:Tissue Scaffolds / Q000737:chemistry*; D014407:Tumor Cells, Cultured / Q000166:cytology
chemical_list: D001672:Biocompatible Materials; D010455:Peptides; D011094:Polyethyleneimine; D004549:Elastin
keywords: 3D cell culture; amide coupling; cell morphology; elastin-like polypeptide; liver tissue model; spheroids