Generation of Multilayered 3D Structures of HepG2 Cells Using a Bio-printing Technique

Abstract

Background/AimsChronic liver disease is a major widespread cause of death, and whole liver transplantation is the only definitive treatment for patients with end-stage liver diseases. However, many problems, including donor shortage, surgical complications and cost, hinder their usage. Recently, tissue-engineering technology provided a potential breakthrough for solving these problems. Three-dimensional (3D) printing technology has been used to mimic tissues and organs suitable for transplantation, but applications for the liver have been rare.MethodsA 3D bioprinting system was used to construct 3D printed hepatic structures using alginate. HepG2 cells were cultured on these 3D structures for 3 weeks and examined by fluorescence microscopy, histology and immunohistochemistry. The expression of liver-specific markers was quantified on days 1, 7, 14, and 21.ResultsThe cells grew well on the alginate scaffold, and liver-specific gene expression increased. The cells grew more extensively in 3D culture than two-dimensional culture and exhibited better structural aspects of the liver, indicating that the 3D bioprinting method recapitulates the liver architecture.ConclusionsThe 3D bioprinting of hepatic structures appears feasible. This technology may become a major tool and provide a bridge between basic science and the clinical challenges for regenerative medicine of the liver.

Liver tissue engineering: From implantable tissue to whole organ engineering

“…Different manufacturing techniques have been used to 3D print hepatic‐like structures, including biomimicry (i.e., identical reproduction of the cellular and extracellular components of a tissue or organ) and minitissue building blocks (i.e., cell sphere assembled in a more complex 3D structure) 27. Cells of the hepatic cell line Hepg2 were printed with alginate as the crosslinking agent, but cell viability was reduced when a high‐extrusion pressure from the printing device was applied 28. Because alginate is characteristically bioinert to mammalian cells and therefore not supportive of cell differentiation and survival, other biomaterials have been explored 29.…”

Reconstruction of structure and function in tissue engineering of solid organs: Toward simulation of natural development based on decellularization

“…Alginate was used to produce hepatic structure through a 3D bioprinting system. Liver architecture was recapitulated via seeding HepG2 cells onto the structure (Jeon et al, ). Besides, chitosan scaffolds were seeded with rat neonatal cardiac cells, which produced a bioengineered open ventricle with high retention rate, desirable biopotential output, and localized cell clusters (Patel, Mohamed, Yazdi, Tasciotti, & Birla, ).…”

3D Culture System for Liver Tissue Mimicking Hepatic Plates for Improvement of Human Hepatocyte (C3A) Function and Polarity

“…Recent studies using in vitro 3D hepatocyte culture include systems based on microcarriers [3], hollow fibers [4][5][6], decellularized liver scaffolds [7][8][9], spheroidal aggregates [10][11][12][13], cell sheets [14][15][16], microfluidic chips [17][18][19][20], and 3D bioprinted cultures [21][22][23][24]. Despite the advantages of increased hepatocyte viability compared to 2D cultures, such systems share a common shortcoming.…”