Precise stacking of decellularized extracellular matrix based 3D cell-laden constructs by a 3D cell printing system equipped with heating modules

Geunseon Ahn,Jun-Young Kim,Songwan Jin,Won-Soo Yun,Dong-Woo Cho,Kyung-Hyun Min,Changhwan Kim,Joo-Yun Won,Jin-Hyung Shim,Jeong-Seok Lee,Donggu Kang

doi:10.1038/s41598-017-09201-5

Abstract

Three-dimensional (3D) cell printing systems allow the controlled and precise deposition of multiple cells in 3D constructs. Hydrogel materials have been used extensively as printable bioinks owing to their ability to safely encapsulate living cells. However, hydrogel-based bioinks have drawbacks for cell printing, e.g. inappropriate crosslinking and liquid-like rheological properties, which hinder precise 3D shaping. Therefore, in this study, we investigated the influence of various factors (e.g. bioink concentration, viscosity, and extent of crosslinking) on cell printing and established a new 3D cell printing system equipped with heating modules for the precise stacking of decellularized extracellular matrix (dECM)-based 3D cell-laden constructs. Because the pH-adjusted bioink isolated from native tissue is safely gelled at 37 °C, our heating system facilitated the precise stacking of dECM bioinks by enabling simultaneous gelation during printing. We observed greater printability compared with that of a non-heating system. These results were confirmed by mechanical testing and 3D construct stacking analyses. We also confirmed that our heating system did not elicit negative effects, such as cell death, in the printed cells. Conclusively, these results hold promise for the application of 3D bioprinting to tissue engineering and drug development.

Highlights

Three-dimensional (3D) cell printing systems have been developed to generate artificial tissues or organs in the field of tissue engineering[1,2,3,4,5,6,7]
We demonstrated that a 3D cell printing system equipped with a heating system dramatically improved the printing fidelity of 3D constructs consisting of decellularized extracellular matrix (dECM) bioink with no cytotoxicity
Skin-derived dECM was produced by decellularization and solubilization processes from native porcine-derived skin tissues (Fig. 1(A))

Summary

Introduction

Three-dimensional (3D) cell printing systems have been developed to generate artificial tissues or organs in the field of tissue engineering[1,2,3,4,5,6,7]. Bioinks with low viscosity have liquid-like properties, resulting in the collapse of 3D-printed constructs in a layer-by-layer process[19, 20, 22, 36,37,38]. As cellular activities, such as proliferation and differentiation, in bioinks are critical to the maturation of artificial tissues or organs[20], suitable rheological properties (i.e. low viscosity) directly related to the cellular microenvironment should be considered during the 3D cell printing process. The printing accuracy and structural integrity of 3D-printed constructs are important for the successful fabrication of 3D tissue analogues during long-term culture[20, 22, 36]

Objectives

Methods

Results

Conclusion