Differentiation of human-induced pluripotent stem cell under flow conditions to mature hepatocytes for liver tissue engineering

Abstract

Hepatic differentiation of human‐induced pluripotent stem cells (hiPSCs) under flow conditions in a 3D scaffold is expected to be a major step forward for construction of bioartificial livers. The aims of this study were to induce hepatic differentiation of hiPSCs under perfusion conditions and to perform functional comparisons with fresh human precision‐cut liver slices (hPCLS), an excellent benchmark for the human liver in vivo. The majority of the mRNA expression of CYP isoenzymes and transporters and the tested CYP activities, Phase II metabolism, and albumin, urea, and bile acid synthesis in the hiPSC‐derived cells reached values that overlap those of hPCLS, which indicates a higher degree of hepatic differentiation than observed until now. Differentiation under flow compared with static conditions had a strong inducing effect on Phase II metabolism and suppressed AFP expression but resulted in slightly lower activity of some of the Phase I metabolism enzymes. Gene expression data indicate that hiPSCs differentiated into both hepatic and biliary directions. In conclusion, the hiPSC differentiated under flow conditions towards hepatocytes express a wide spectrum of liver functions at levels comparable with hPCLS indicating excellent future perspectives for the development of a bioartificial liver system for toxicity testing or as liver support device for patients.

Improving functional re-endothelialization of acellular liver scaffold using REDV cell-binding domain

“…However, stem cells derived from current methods retain epigenetic memory, which could impair their potential to differentiate into cells of all lineages by predisposing iPS cells to differentiate more readily into their parental cells than others [53] [54]. Nevertheless, the use of iPS for liver engineering represent an important direction for research and further studies are in need to obtain scalable and efficient methods to differentiate pluripotent cells to liver cells equivalent [55] [56] [57].…”

<p>Graphene-based 3D scaffolds in tissue engineering: fabrication, applications, and future scope in liver tissue engineering</p>

“…The biochemical and histopathological analysis reported serum biochemical changes, escalation in the level of ROS, liver enzymes (alanine, aspartate, alkaline phosphates), concentration of lipid hydroperoxide and damage to liver tissue 265. Advances in stem cell-based therapies, liver-on-a-chip technologies, disease modeling, drug testing, and organ-specific vascularization technique offer promise to the liver regenerative therapy 266–268. In addition, optimization of graphene-based compatible 3D scaffolds along with stem cell sources and bioengineering approaches could deliver a fully functional transplantable liver for the future generations.…”

Human hepatocyte systems for in vitro toxicology analysis

“…A hallmark of pluripotent stem cells is their intrinsic proliferation capacity and their potential to divide into cell types of all three germ layers. It is thus not surprising that there have been numerous attempts to generate functional hepatocytes from embryonic (ESCs) and induced pluripotent stem cells (iPS) [58][59][60][61][62]. ESCs represent a natural pluripotent stem cell type and their differentiation potential is thought to be superior to iPS cells [63].…”