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Isolation of rat hepatocytes
Peggy Papeleu1, Tamara Vanhaecke, Tom Henkens
1Department of Toxicology, Vrije Universiteit Brussel, Belgium.
Methods in Molecular Biology (Clifton, N.J.)
|May 25, 2006
Summary
This study details a modified collagenase perfusion method for high-yield isolation of intact primary rat hepatocytes. A new method using histone deacetylase inhibition prevents dedifferentiation in cultured hepatocytes.
Area of Science:
- Hepatology
- Drug Development
- Cell Biology
Background:
- In vitro liver models are crucial for preclinical drug development.
- High-yield preparation of intact hepatocytes is a key advancement for cell models.
- Hepatocyte dedifferentiation during culture is a significant challenge.
Purpose of the Study:
- To outline practical aspects of a modified two-step collagenase perfusion method for rat liver hepatocyte isolation.
- To provide a troubleshooting guide for successful primary rat hepatocyte isolation.
- To present a novel method for inhibiting hepatocyte dedifferentiation.
Main Methods:
- Modified two-step collagenase perfusion technique (Seglen's original procedure).
- Application to rat liver, with notes on applicability to other species.
- Histone deacetylase (HDAC) inhibition to prevent cell-cycle reentry.
Main Results:
- Successful high-yield preparation of intact primary rat hepatocytes.
- Identification of critical parameters for optimal hepatocyte isolation.
- Demonstration of HDAC inhibition's ability to prevent dedifferentiation.
Conclusions:
- The modified collagenase perfusion method is effective for isolating high-quality primary rat hepatocytes.
- Inhibiting histone deacetylase activity offers a promising strategy to maintain hepatocyte function in culture.
- These advancements improve the utility of in vitro liver models for drug development.