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

Abstract

In vitro 3D hepatocyte culture constitutes a core aspect of liver tissue engineering. However, conventional 3D cultures are unable to maintain hepatocyte polarity, functional phenotype, or viability. Here, we employed microfluidic chip technology combined with natural alginate hydrogels to construct 3D liver tissues mimicking hepatic plates. We comprehensively evaluated cultured hepatocyte viability, function, and polarity. Transcriptome sequencing was used to analyze changes in hepatocyte polarity pathways. The data indicate that, as culture duration increases, the viability, function, polarity, mRNA expression, and ultrastructure of the hepatic plate mimetic 3D hepatocytes are enhanced. Furthermore, hepatic plate mimetic 3D cultures can promote changes in the bile secretion pathway via effector mechanisms associated with nuclear receptors, bile uptake, and efflux transporters. This study provides a scientific basis and strong evidence for the physiological structures of bionic livers prepared using 3D cultures. The systems and cultured liver tissues described here may serve as a better in vitro 3D culture platform and basic unit for varied applications, including drug development, hepatocyte polarity research, bioartificial liver bioreactor design, and tissue and organ construction for liver tissue engineering or cholestatic liver injury.

Organ-on-a-Chip systems for new drugs development

“…The OoC can reproduce particular conditions, such as the presence of specific pathologies, age and sex differences, or metabolic modifications [3,7,8,15,[42][43][44]. The ability of OoC systems to mimic the functionality of an organ allows to visualize the impact of a condition on the organ's function.…”

“…Jia et al described a 3D hepatic system for liver injury studies by controlling hepatocyte viability, function, morphology and polarity, and thus, maintaining the tissue morphology, mRNA expression, bile secretion and a better cell viability compared to other types of cell cultures. They controlled the morphology encapsulating the cell culture inside a alginate hydrogel (figure 8A) [43]. Freyer et al built an OoC model for hepatotoxicity testing, highlighting the importance of the control and monitoring of chemical physiological parameters (such as lactate production and ammonia release) (figure 8B).…”

“…plate structures, and a ii) system of syringe pumps supplying the microfluidic channels of the OoC with solutions. Adapted from [43] under terms of the Creative Commons Attribution License. Copyright (2020).…”

Harnessing Inorganic Nanoparticles to Direct Macrophage Polarization for Skeletal Muscle Regeneration

“…It is therefore evident that the macrophage response induced by scaffolds (both natural and synthetic) is highly variable, mainly because there are many factors involved (biomaterial, composition, topography, porosity, etc.). Furthermore, it is arduous to recreate the complexity of the microenvironmental stimuli naturally polarizing macrophages; thus, novel approaches such as microfluidic systems are trying to recreate in vitro, the in vivo stimulation [ 40 ].…”

Argininosuccinate synthase 1 suppresses tumor progression through activation of PERK/eIF2α/ATF4/CHOP axis in hepatocellular carcinoma

“…In contrast, a complex three-dimensional (3D) cell culture system better simulates cellular context and the therapeutically relevant parameters of the in vivo TME, such as pH, oxygen level, metabolite gradients, growth factor penetration, and distribution of proliferating/necrotic cells [ 8 , 9 ]. In particular, liver cells in a 3D culture system better recapitulate numerous physiological liver functions, including albumin and urea synthesis, bile secretion, and cell polarization [ 10 , 11 ].…”