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Updated: Dec 3, 2025

An Improved Time- and Labor- Efficient Protocol for Mouse Primary Hepatocyte Isolation

An Improved Time- and Labor- Efficient Protocol for Mouse Primary Hepatocyte Isolation

Published on: October 25, 2021

Protocol for Primary Mouse Hepatocyte Isolation.

Meital Charni-Natan¹, Ido Goldstein¹

¹Institute of Biochemistry, Food Science and Nutrition, The Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, POB 12, Rehovot 7610001, Israel.

|October 28, 2020

Summary

This summary is machine-generated.

Obtaining high-yield primary mouse hepatocytes is difficult. This study presents an optimized collagenase perfusion protocol to improve hepatocyte yield and viability for liver research.

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Isolation and Characterization of Mouse Primary Liver Sinusoidal Endothelial Cells

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Published on: December 16, 2021

Area of Science:

Hepatology and cell biology research.
Biomedical research utilizing *ex vivo* models.

Background:

Primary hepatocytes are crucial for studying liver physiology *ex vivo*.
Current methods for isolating primary mouse hepatocytes often yield insufficient numbers of viable cells, hindering research.
Technical challenges limit the widespread use of primary hepatocytes.

Purpose of the Study:

To present an optimized protocol for obtaining high yields of viable primary mouse hepatocytes.
To improve upon the classic two-step collagenase perfusion technique.
To reduce procedure duration and enhance cell yield and viability.

Main Methods:

Utilized a two-step collagenase perfusion technique for liver dissociation.
Optimized liver washing and hepatocyte separation procedures.
Standardized cell culture methods post-isolation.

Main Results:

Achieved significantly reduced procedure duration compared to standard methods.
Demonstrated improved yields of viable primary mouse hepatocytes.
Enhanced overall quality and usability of isolated hepatocytes for research.

Conclusions:

The optimized protocol effectively addresses the challenge of low yield and viability in primary hepatocyte isolation.
This improved technique facilitates more robust and reproducible *ex vivo* studies of liver function.
Enables broader application of primary mouse hepatocytes in biomedical research.