title: Three-dimensional growth of human hepatoma C3A cells within alginate beads for fluidized bioartificial liver.
abstract: OBJECTIVE
  Alginate beads allow cultivation of cells in a 3-dimensional environment. The aim of our study was to assess the influence of a 3-dimensional culture in alginate microbeads, on hepatic cell metabolism.
  
  
  METHODS
  We used 2 types of alginate: low viscosity (LV) and medium viscosity (MV). The hepatic cell line C3A was encapsulated in alginate beads. Cells were cultured for 2 weeks. Using scanning electron microscopy, the morphology of 3D structures and the surfaces of cells were analyzed. Fluidized bed bioartificial liver experiments were performed 24 hours, 7, and 14 days after bead formation.
  
  
  RESULTS
  Two different cell growth types in alginate beads were observed: channel-like structures and spherical aggregates characteristic of LV and MV alginate, respectively. A significant increase in albumin synthesis was observed in long-term culture. Formation of characteristic hepatic cell microvilli on cell surfaces was observed under scanning electron microscopy for both types of alginate. Prolonged static cultivation of C3A cells within the alginate beads in both types of alginates caused significant increases in albumin production in the fluidized bioreactor.
  
  
  CONCLUSIONS
  Cultivation of the hepatic C3A cells within the alginate microbeads significantly improved bioreactor effectiveness in albumin production. The presence of extensions of cell membranes on the surface of hepatoma cells in 3-dimensional culture within the alginate beads indicated formation of microvilli-like structures characteristic of normal hepatocytes.
authors.affiliation: Institute of Biocybernetics and Biomedical Engineering, Polish Academy of Sciences, Warsaw, Poland. kinas@ibib.waw.pl
authors.forename: A
authors.initials: A
authors.lastname: Kinasiewicz
authors.forename: A
authors.initials: A
authors.lastname: Gautier
authors.forename: D
authors.initials: D
authors.lastname: Lewiska
authors.forename: A
authors.initials: A
authors.lastname: Smietanka
authors.forename: C
authors.initials: C
authors.lastname: Legallais
authors.forename: A
authors.initials: A
authors.lastname: Weryński
mesh_terms: D000418:Albumins / Q000378:metabolism; D000464:Alginates / Q000737:chemistry*; D001672:Biocompatible Materials; D019149:Bioreactors; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D018929:Cell Culture Techniques; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D020723:Glucuronic Acid / Q000737:chemistry; D006603:Hexuronic Acids / Q000737:chemistry; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism / Q000473:pathology* / Q000648:ultrastructure; D019164:Liver, Artificial; D008855:Microscopy, Electron, Scanning; D008863:Microspheres; D018874:Spheroids, Cellular; D013997:Time Factors; D015854:Up-Regulation; D014783:Viscosity
chemical_list: D000418:Albumins; D000464:Alginates; D001672:Biocompatible Materials; D006603:Hexuronic Acids; D020723:Glucuronic Acid