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In vitro metabolic zonation through oxygen gradient on a chip
Federica Tonon1, Giovanni Giuseppe Giobbe2,3, Alessandro Zambon3
1Dept. of Life Sciences, University of Trieste, Trieste, 34127, Italy.
Scientific Reports
|September 21, 2019
Summary
An oxygen gradient in a microfluidic device successfully induced metabolic zonation in differentiating human embryonic stem cells (hESCs) into hepatocytes. This controlled oxygen environment mimics in vivo conditions, enabling functional metabolic zonation and offering insights into liver physiology and disease.
Area of Science:
- * Stem cell biology
- * Liver biology
- * Bioengineering
Background:
- * Hepatic zonation, the spatial organization of metabolic functions within the liver lobule, is crucial for liver homeostasis.
- * Oxygen gradients across the hepatic lobule are known to influence hepatocyte gene expression and metabolism.
- * Understanding the role of oxygen in establishing hepatic zonation is vital for both physiological and pathological contexts.
Purpose of the Study:
- * To investigate if a controlled oxygen gradient, generated in vitro using a microfluidic device, is sufficient to induce functional metabolic zonation during human embryonic stem cell (hESC)-derived hepatocyte differentiation.
- * To mimic in vivo hepatic zonation conditions in an in vitro model for studying liver cell development and function.
Main Methods:
- * Design and utilization of a microfluidic device to generate a stable oxygen gradient.
- * Differentiation of hESCs from endoderm towards hepatocytes under the applied oxygen gradient.
- * Characterization of cell differentiation stages using specific markers (SOX-17, AFP, HNF-4α, ALB).
- * Assessment of metabolic zonation through glycogen storage (PAS staining), CYP3A4 expression, and quantitative real-time PCR for zonated genes (GLUL, CTNNB, CCND1, CPS1, MKI67, CCNA).
Main Results:
- * The oxygen gradient successfully induced zonated glycogen storage, with higher accumulation in hepatocytes under high oxygen (high pO2).
- * Genes associated with low oxygen conditions (GLUL, CTNNB, CCND1) showed higher expression in low pO2 regions.
- * Genes associated with high oxygen conditions (CPS1, ALB, MKI67, CCNA) exhibited significantly higher expression in high pO2 regions.
- * This demonstrates the establishment of functional metabolic zonation in vitro.
Conclusions:
- * The developed microfluidic device and oxygen gradient effectively instruct the establishment of functional metabolic zonation in differentiating hESCs.
- * This in vitro model provides a powerful tool to study hepatocyte physiology and pathology in relation to oxygen levels.
- * Future research can leverage this system to deepen understanding of liver function and disease mechanisms influenced by oxygen gradients.
