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Quantitative and Temporal Control of Oxygen Microenvironment at the Single Islet Level
Published on: November 17, 2013
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An in vitro hepatic zonation model with a continuous oxygen gradient in a microdevice.
Asako Sato1, Kanae Kadokura2, Hideyuki Uchida1
1Graduate School of Fundamental Science and Technology, Keio University, 3-14-1 Hiyoshi, Kouhoku-ku, Yokohama, Kanagawa 223-8522, Japan.
Biochemical and Biophysical Research Communications
|December 3, 2014
Summary
Researchers developed a microdevice to mimic the liver
Area of Science:
- * Biomedical Engineering
- * Cell Biology
- * Organ-on-a-chip Technology
Background:
- * Hepatic lobules exhibit functional zonation due to regional differences in metabolic enzyme expression.
- * Oxygen gradients are critical for hepatic zonation, but replicating this microenvironment in vitro is challenging.
- * Conventional cell cultures lack the necessary oxygen gradients for accurate hepatocyte studies.
Purpose of the Study:
- * To design and validate a novel microdevice capable of recreating the oxygen gradient found in hepatic lobules.
- * To enable long-term culture of primary hepatocytes under physiologically relevant oxygen conditions.
- * To provide a platform for studying liver zonation and cellular respiration in vitro.
Main Methods:
- * Development of a microfluidic cell culture device with controlled oxygen gradients.
- * Utilization of laser-assisted phosphorescence quenching for real-time oxygen gradient monitoring.
- * Continuous medium exchange to maintain stable oxygen gradients during culture.
- * Measurement of hepatocyte oxygen consumption rates at different oxygen partial pressures.
- * Gene expression analysis (RT-PCR) to assess metabolic zonation markers.
Main Results:
- * The microdevice successfully established and maintained stable oxygen gradients mimicking periportal (PP) and pericentral (PC) regions.
- * Hepatocyte oxygen consumption rates were quantified under simulated PP (70.0 mmHg) and PC (31.4 mmHg) conditions.
- * The microdevice demonstrated sensitivity to cellular respiration changes, as shown by Antimycin A treatment.
- * Gene expression analysis revealed distinct upregulation of PEPCK (PP) and GK (PC) in the respective micro-regions, confirming functional zonation.
Conclusions:
- * The developed microdevice effectively reproduces the hepatic lobular microenvironment in vitro.
- * This technology allows for precise control of oxygen gradients, crucial for studying organ zonation.
- * The platform holds potential for various organ-on-a-chip applications and modeling liver diseases.
