Multi‐omics analysis of hiPSCs‐derived HLCs matured on‐chip revealed patterns typical of liver regeneration

Abstract

Maturation of human‐induced pluripotent stem cells (hiPSCs)‐derived hepatocytes‐like cells (HLCs) toward a complete hepatocyte phenotype remains a challenge as primitiveness patterns are still commonly observed. In this study, we propose a modified differentiation protocol for those cells which includes a prematuration in Petri dishes and a maturation in microfluidic biochip. For the first time, a large range of biomolecular families has been extracted from the same sample to combine transcriptomic, proteomic, and metabolomic analysis. After integration, these datasets revealed specific molecular patterns and highlighted the hepatic regeneration profile in biochips. Overall, biochips exhibited processes of cell proliferation and inflammation (via TGFB1) coupled with anti‐fibrotic signaling (via angiotensin 1‐7, ATR‐2, and MASR). Moreover, cultures in this condition displayed physiological lipid‐carbohydrate homeostasis (notably via PPAR, cholesterol metabolism, and bile synthesis) coupled with cell respiration through advanced oxidative phosphorylation (through the overexpression of proteins from the third and fourth complex). The results presented provide an original overview of the complex mechanisms involved in liver regeneration using an advanced in vitro organ‐on‐chip technology.

Microfluidic devices: The application in TME modeling and the potential in immunotherapy optimization

“…Different from traditional 2D cell culture models, organ chips with microchannels allowed the fluids to flow across the cell chambers, which enabled the recapitulation for in vivo physical conditions such as vascular perfusion, air-liquid interfaces, shear stresses as well as the physical and chemical gradients ( Murugesan et al, 2017 ; Liu et al, 2021 ). For instance, microfluidic chips were used to culture the human‐induced pluripotent stem cells (hiPSCs)‐derived hepatocytes-like cells (HLCs) ( Danoy et al, 2021 ). And the multi‐omics analysis of the chip demonstrated a typical signature of a liver regenerative process, which provided an original overview of the sophisticated mechanisms of liver regeneration by the use of microfluidic technology ( Danoy et al, 2021 ).…”

“…For instance, microfluidic chips were used to culture the human‐induced pluripotent stem cells (hiPSCs)‐derived hepatocytes-like cells (HLCs) ( Danoy et al, 2021 ). And the multi‐omics analysis of the chip demonstrated a typical signature of a liver regenerative process, which provided an original overview of the sophisticated mechanisms of liver regeneration by the use of microfluidic technology ( Danoy et al, 2021 ). In addition, some sophisticated organ chips even succeed in modeling organ-relevant mechanical activity by manipulating the organotypic tissue interfaces with the designed hollow side chambers ( Choi et al, 2015 ; Hassell et al, 2017 ).…”

Mass spectrometry-based quantitation combined with time-dependent metabolomics to discover metabolic features in human neurogenesis using neural constructs generated from neural progenitor cells

“…11,12 Metabolomics offers the closest direct measurement of a cell's physiological activity, and has advanced efforts to characterize cells' fate, identify biomarkers, and investigate metabolic pathways. 13 Recently, analyses of the metabolic changes accompanying maturation of several human in vitro models, such as the liver 14,15 and heart, 16 have been performed to better characterize and understand these organs. However, metabolomics study on human neurodevelopment is greatly lacking.…”

Development of Liver-on-Chip Integrating a Hydroscaffold Mimicking the Liver’s Extracellular Matrix

“…In previous works, our group has developed liver-on-chip models with different hepatic cells (HepG2/C3A, HepaRG, PHH, primary rat hepatocytes and human induced pluripotent stem cells hiPSCs) to investigate liver metabolism [ 27 , 28 ], drugs and pesticide toxicity [ 29 , 30 , 31 ], and liver regeneration and the development process [ 32 ]. Recently, we integrated an alginate cryogel into our biochip to promote 3D cell organization.…”