Fabrication of 3D-culture platform with sandwich architecture for preserving liver-specific functions of hepatocytes using 3D bioprinter

J Biomed Mater Res A. 2017 Jun;105(6):1583-1592. doi: 10.1002/jbm.a.35905. Epub 2017 Mar 23.

Abstract

The development of new three-dimensional (3D) cell culture system that maintains the physiologically relevant signals of hepatocytes is essential in drug discovery and tissue engineering research. Conventional two-dimensional (2D) culture yields cell growth, proliferation, and differentiation. However, gene expression and signaling profiles can be different from in vivo environment. Here, we report the fabrication of a 3D culture system using an artificial scaffold and our custom-made inkjet 3D bioprinter as a new strategy for studying liver-specific functions of hepatocytes. We built a 3D culture platform for hepatocytes-attachment and formation of cell monolayer by interacting the galactose chain of galactosylated alginate gel (GA-gel) with asialoglycoprotein receptor (ASGPR) of hepatocytes. The 3D geometrical arrangement of cells was controlled by using 3D bioprinter, and cell polarity was controlled with the galactosylated hydrogels. The fabricated GA-gel was able to successfully promote adhesion of hepatocytes. To observe liver-specific functions and to mimic hepatic cord, an additional parallel layer of hepatocytes was generated using two gel sheets. These results indicated that GA-gel biomimetic matrices can be used as a 3D culture system that could be effective for the engineering of liver tissues. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part A: 105A: 1583-1592, 2017.

Keywords: 3D bioprinter; 3D culture; galactosylated alginate (GA-gel); hepatocyte; tissue engineering.

MeSH terms

  • Alginates / chemistry
  • Alginates / metabolism*
  • Animals
  • Asialoglycoprotein Receptor / metabolism*
  • Biocompatible Materials / chemistry
  • Biocompatible Materials / metabolism*
  • Cell Adhesion
  • Cells, Cultured
  • Equipment Design
  • Galactose / analogs & derivatives
  • Galactose / metabolism*
  • Glucuronic Acid / chemistry
  • Glucuronic Acid / metabolism
  • Hepatocytes / cytology*
  • Hepatocytes / metabolism
  • Hexuronic Acids / chemistry
  • Hexuronic Acids / metabolism
  • Mice, Inbred ICR
  • Printing, Three-Dimensional*
  • Tissue Engineering / instrumentation*

Substances

  • Alginates
  • Asialoglycoprotein Receptor
  • Biocompatible Materials
  • Hexuronic Acids
  • Glucuronic Acid
  • Galactose