Transcriptomic and proteomic studies suggest the establishment of advanced zonation-like profiles in hiPSCs-derived LSECs and CPM-positive LPCs cocultured in a fluidic microenvironment.

AIM: Hepatic zonation is a physiological feature of the liver, known to be key in the regulation of the metabolism of nutrients and xenobiotics and the biotransformation of numerous substances. However, the reproduction of this phenomenon remains challenging in vitro as only part of the processes involved in the orchestration and maintenance of zonation are fully understood. The recent advances in organ-on-chip technologies, which allow for the integration of multicellular 3D tissues in a dynamic microenvironment could offer solutions for the reproduction of zonation within a single culture vessel.

METHODS: An in-depth analysis of zonation-related mechanisms observed during the coculture of hiPSCs-derived CPM-positive liver progenitor cells and hiPSCs-derived liver sinusoidal endothelial cells within a microfluidic biochip was performed.

RESULTS: Hepatic phenotypes were confirmed in terms of albumin secretion, glycogen storage, CYP450 activity, and expression of specific endothelial markers such as PECAM-1, RAB5A and CD109. Further characterization of the patterns observed in the comparison of the transcription factors motifs activities, the transcriptomic signature, and the proteomic profile expressed at the inlet and the outlet of the microfluidic biochip confirmed the presence of zonation-like phenomena within the biochips. Especially, differences related to Wnt/β-catenin, TGFβ, mTOR, HIF1, and AMPK signaling, to the metabolism of lipids, and cellular remolding were observed.

CONCLUSIONS: The present study shows the interest in combining coculture of hiPSCs-derived cellular models and microfluidic technologies for reproducing in vitro complex mechanisms such as liver zonation and further incites the use of those solutions for accurate reproduction of in vivo situations. This article is protected by copyright. All rights reserved.