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

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.

Tissue Engineering in Liver Regenerative Medicine: Insights into Novel Translational Technologies

“…A practical biomaterial for 3D bio-printing is usually a biocompatible substance, which should be easily manipulated and it could maintain or even enhance cell viability and functions [70]. Different types of 3D bio-printing technologies have been introduced so far, including ink-jet-based bio-printing [71], laser-assisted bio-printing [72], extrusion-based bio-printing [73], stereo-lithography-based bio-printing [74] and microvalve-based bio-printing [75] and many other novel emerging technologies [76] (Table 2). Among these technologies, probably extrusion-based bio-printing has been the most widely used one to construct living 3D tissues and organs [77].…”

“…In conclusion, 3D bio-printing is a promising technology in the field of bio-artificial organ generation, which may overcome various limitations encountered in different models [66] and improve maturation of hepatocyte like cells (HLCs) [75]. Furthermore, this technology could preserve ex vivo hepatocyte function and maintenance [71]. Also, thanks to the multi-nozzle 3D bio-printers and novel biocompatible polymers, the artificial organs could be more similar to the original tissue compartments [77].…”

“…Early personalized drug screening and liver pathophysiology studies in vitro [74] Inkjet-based Galactosylated alginate gel (GA-gel) Mouse primary hepatocyte preservation of functions and polarity in hepatocytes [71] Microvalve-based Alginate hPSC…”

Bioprinting for Liver Transplantation

“…Arai et al [73] used an inkjet 3D bioprinter to fabricate a 3D culture system using an artificial scaffold for studying the liver-specific functions of hepatocytes. The printed construct expressed liver-specific proteins and receptors such as albumin, MPR2, and asialoglycoprotein receptor (ASGPR), thus proving the functionality of the printed liver tissue.…”

Vascularized cardiac tissue construction with orientation by layer-by-layer method and 3D printer

“…However, reports about fabrication of 3D cardiac tissue with an oriented and vascularized structure are seldom mentioned and have many problems such as the lack of bottom-up technology. In order to solve this problem, the 3D printer technology which precisely controls the placement of cells and materials is attracting great attention [31][32][33][34][35][36] .…”