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Updated: May 17, 2026
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Protocol for Isolation of Primary Human Hepatocytes and Corresponding Major Populations of Non-parenchymal Liver Cells
Published on: March 30, 2016
Preparation of hepatocytes
1XenoTech, LLC, Kansas City, Kansas, USA.
Current Protocols in Toxicology
|October 10, 2012
Summary
This study details a method for isolating intact, viable hepatocytes for drug metabolism research. The protocol covers liver tissue digestion, perfusion techniques, and cryopreservation for long-term storage and integrated studies.
Area of Science:
- Pharmacology and Toxicology
- Cell Biology
- Biochemistry
Background:
- Isolated hepatocytes are crucial for in vitro xenobiotic metabolism studies.
- Established methods are needed for reliable hepatocyte isolation and preservation.
- Drug metabolism research relies on high-quality, viable cellular models.
Purpose of the Study:
- To describe a robust process for isolating intact, viable hepatocytes.
- To provide methods for both in situ and ex vivo liver perfusion.
- To outline cryopreservation techniques for long-term hepatocyte storage.
Main Methods:
- Two-step enzymatic digestion of liver tissue (animal and human).
- In situ liver perfusion and perfusion of excised liver tissue.
- Cryopreservation protocols for isolated hepatocytes.
Main Results:
- Successful isolation of intact, viable hepatocytes.
- Demonstration of two distinct perfusion methodologies.
- Established cryopreservation enabling long-term storage.
Conclusions:
- The described protocol enables the isolation and preservation of hepatocytes for integrated drug metabolism studies.
- This method supports in vitro xenobiotic metabolism research using reliable cellular models.
- Cryopreserved hepatocytes offer a valuable resource for ongoing drug metabolism investigations.
