Bioprinting of a Hepatic Tissue Model Using HumanInduced Pluripotent Stem Cell-derived Hepatocytes for Drug-Induced Hepatotoxicity Evaluation

Abstract

Three-dimensional (3D) bioprinting technology is an effective method for exploring the biological functions of hepatocytes by building biomimetic 3D microenvironments. Various hepatic tissue models have been developed for disease modeling, drug screening, and tissue regeneration using 3D bioprinting technology. Human-induced pluripotent stem cells (hiPSCs) are a promising cell source for the generation of functional hepatocytes for bioprinting. In this study, we introduced hiPSC-derived hepatocytes (hiPSC-Heps) as mature hepatocytes for the bioprinting of a 3D hepatic tissue model. The 3D-printed (3DP) model facilitated the formation of hiPSC-Hep spheroids with higher viability and proliferation than the commonly used non-printed sandwich-cultured model. hiPSC-Heps in the 3DP model exhibited higher mRNA expression of liver-specific functions than those in the two-dimensional-cultured model. Moreover, enhanced secretion of liver function-related proteins, including α-1-antitrypsin, albumin, and blood urea nitrogen, was observed in the 3DP model. For the evaluation of acetaminophen-induced hepatotoxicity, the 3DP model exhibited a favorable drug response with upregulation of the drug metabolism-related gene cytochrome P450-1A2 (CYP1A2). Overall, the bioprinted hepatic tissue model showed great biofunctional and drug-responsive performance, which could be potentially applied in in vitro toxicological studies.

iPSC-derived cells for whole liver bioengineering

“…Biofabrication, defined as the automated production of functional tissues and organs, is a burgeoning field ( Ambhorkar et al, 2020 ). Within the area, three-dimensional (3D) bioprinting holds promise for the manufacturing of artificial organs, enabling not only precise cell deposition, but shape and size control combining biocompatible bioinks and bioprinting techniques ( He et al, 2022 ).…”

Toward 3D‐Bioprinted Models of the Liver to Boost Drug Development

“…[83] Currently, the most diffuse type of hepatic macrophages for 3D-bioprinting applications are Kupffer-like cells, derived from human iPSCs. [70,84] However, Kupffer and Kupffer-like cells are not the only option to introduce immune cells into hepatic models. As a matter of fact, the exploitation of primary human peripheral blood monocytes for the biofabrication of liver in vitro models was also recently documented.…”

Engineered 3D liver-tissue model with minispheroids formed by a bioprinting process supported with in situ electrical stimulation

“…For in vitro liver tissue fabrication, a bioprinting technique has been employed to build hepatic models, with a particular focus on mimicking the liver tissue-specific unit of the hexagonal-structured hepatic lobule and providing vessel formation [ 12 , 13 ]. Table 1 provides an overview of the previous studies on bioprinted liver models [ [14] , [15] , [16] , [17] , [18] , [19] , [20] , [21] ]. Particularly, X. Ma et al [ 19 ].…”

“… Method Cells Bioink Strategies Key results Ref. Extrusion-based bioprinting L02 Collagen-chitosan Bioprinting cell-laden liver construct with a grid pattern Addition of hepatocyte growth factor Improvement in liver tissue restoration and function in vivo Lacks in patterning hepatic lobule Lacks in using multiple cell types [ 14 ] Extrusion-based bioprinting Huh7 GelMA Comprising triple cells by seeding HUVECs onto the HepaRG/LX-2 mixture-laden structure Treatment of TGF-b1 for fibrosis Improvement in hepatic cell function Enabling the observation of collagen deposition enabling the fabrication of relevant pathological model Lacks in patterning hepatic lobule [ 15 ] HepaRG HUVEC LX-2 Extrusion-based bioprinting hiPSC-Hep Alginate-gelatin Using a promising cell source for the generation of functional hepatocytes Formation of hepatocyte spheroid by bioprinting Improvement in liver tissue function Exhibiting favorable drug response Lacks in patterning hepatic lobule Lacks in using multiple cell types [ 16 ] Extrusion-based core-shell bioprinting HepG2 Alginate-MC Using two bioinks : HUVEC/NHDF-laden collagen-based bioink : HepG2-laden alginate-based bioink Using core (HUVEC/NHDF)-shell (HepG2) nozzle to separate cell region Addition of human blood plasma into the shell bioink …”