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Alkoxyresorufin O-dealkylase assay using a rat hepatocyte spheroid microarray
Yusuke Sakai1, Tomoko Tanaka, Junji Fukuda
1Department of Life and Environment Engineering, The University of Kitakyushu, 1-1 Hibikino, Wakamatsu-ku, Kitakyushu, Fukuoka 808-0135, Japan.
Journal of Bioscience and Bioengineering
|March 16, 2010
Summary
Hepatocyte spheroid microarrays (HSM) show enhanced drug metabolism compared to traditional cell cultures. This advanced platform maintains liver function for over two weeks, offering a promising tool for drug testing.
Area of Science:
- Hepatology
- Drug Metabolism
- Biotechnology
Background:
- Hepatocyte spheroids maintain liver functions, making them suitable for cell-based assays.
- Traditional cell culture methods may not fully replicate in vivo liver metabolic capabilities.
- There is a need for robust platforms to accurately assess drug metabolism.
Purpose of the Study:
- To investigate the drug metabolic function of a novel hepatocyte spheroid microarray (HSM) chip.
- To evaluate the response of the HSM chip to known cytochrome P450 (CYP) inducers.
- To assess the long-term stability of liver function in the HSM chip.
Main Methods:
- Primary rat hepatocytes were cultured into spheroids on a microarray chip (HSM).
- An alkoxyresorufin O-dealkylase assay system was employed to measure enzyme activity.
- Hepatocyte spheroid aggregates were exposed to inducers like 3-methylcholanthrene (3-MC) and sodium phenobarbital.
Main Results:
- The HSM chip exhibited 5- to 10-fold higher ethoxyresorufin O-dealkylase (EROD) activity compared to monolayer hepatocytes.
- Enzyme activities (EROD, MROD, BROD, PROD) were induced by specific CYP inducers, correlating with increased gene expression.
- Sustained metabolic function was observed in the HSM chip for at least two weeks.
Conclusions:
- The hepatocyte spheroid microarray (HSM) chip demonstrates superior drug metabolic capacity.
- The HSM chip shows significant responsiveness to CYP inducers, mimicking in vivo responses.
- This platform offers a stable and effective tool for drug metabolic assays, potentially improving drug development screening.
