High density of immobilized galactose ligand enhances hepatocyte attachment and function

J Biomed Mater Res A. 2003 Dec 15;67(4):1093-104. doi: 10.1002/jbm.a.10033.

Abstract

Galactosylated surface is an attractive substrate for hepatocyte culture because of the specific interaction between the galactose ligand and the asialoglycoprotein receptor on hepatocytes. In this study, we described a scheme to achieve high density of immobilized galactose ligands on polyethylene terephthalate (PET) surface by first surface-grafting polyacrylic acid on plasma-pretreated PET film under UV irradiation, followed by conjugation of a galactose derivative (1-O-(6'-aminohexyl)-D-galactopyranoside) to the grafted polyacrylic acid chains. A high galactose density of 513 nmol/cm(2) on the PET surface was used in this study to investigate the behavior of cultured hepatocyte. This engineered substrate showed high affinity to fluorescein isothiocyanate-lectin binding. Primary rat hepatocytes, when seeded at a density of 2 x 10(5) cells/cm(2), attached to the galactosylated PET substrate at a similar efficiency compared with collagen-coated substrate. The hepatocytes spontaneously formed aggregates 1 day after cell seeding and showed better maintenance of albumin secretion and urea synthesis functions than those cultured on collagen-coated surface.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acrylic Resins / chemistry
  • Albumins / metabolism
  • Animals
  • Cell Adhesion / physiology*
  • Cell Culture Techniques / methods*
  • Cell Size
  • Cells, Cultured
  • Fluorescein-5-isothiocyanate / analogs & derivatives
  • Fluorescein-5-isothiocyanate / metabolism
  • Galactose / chemistry*
  • Galactose / metabolism
  • Hepatocytes / physiology*
  • Hepatocytes / ultrastructure
  • Ligands
  • Molecular Structure
  • Plant Lectins / metabolism
  • Polyethylene Terephthalates / chemistry
  • Polyethylene Terephthalates / metabolism
  • Protein Binding
  • Rats
  • Surface Properties
  • Urea / metabolism

Substances

  • Acrylic Resins
  • Albumins
  • Ligands
  • Plant Lectins
  • Polyethylene Terephthalates
  • carbopol 940
  • Urea
  • Fluorescein-5-isothiocyanate
  • Galactose