Protocol for Primary Mouse Hepatocyte Isolation

STAR Protoc. 2020 Aug 13;1(2):100086. doi: 10.1016/j.xpro.2020.100086. eCollection 2020 Sep 18.


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.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Separation / methods*
  • Cells, Cultured
  • Collagenases
  • Hepatocytes / metabolism*
  • Liver / cytology
  • Mice
  • Mice, Inbred C57BL
  • Perfusion
  • Primary Cell Culture / methods*


  • Collagenases

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