Protocol for Primary Mouse Hepatocyte Isolation

Meital Charni-Natan; Ido Goldstein

doi:10.1016/j.xpro.2020.100086

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.

Authors

Meital Charni-Natan¹, Ido Goldstein¹

Affiliation

¹ Institute 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.

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*

Substances

Collagenases

Grants and funding

CIHR/Canada