Protocol for Primary Mouse Hepatocyte Isolation
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Updated: Dec 3, 2025
An Improved Time- and Labor- Efficient Protocol for Mouse Primary Hepatocyte Isolation
Published on: October 25, 2021
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.
Abstract:
Primary hepatocytes are a vital tool in various biomedical research disciplines, serving as an ex vivo model for liver physiology. Obtaining high yields of viable primary mouse hepatocytes is technically challenging, limiting their use. Here, we present an improved protocol based on the classic two-step collagenase perfusion technique. The liver is washed by perfusion, hepatocytes are dissociated by collagenase, separated from other cells, and cultured. This protocol was optimized to significantly reduce procedure duration and improve hepatocyte yield and viability.
