Isolation and primary culture of rat hepatic cells

Ling Shen, Allix Hillebrand, David Q.H. Wang, Min Liu

Research output: Contribution to journalArticlepeer-review

130 Scopus citations


Primary hepatocyte culture is a valuable tool that has been extensively used in basic research of liver function, disease, patho physiology, pharmacology and other related subjects. The method based on two-step collagenase perfusion for isolation of intact hepatocytes was first introduced by Berry and Friend in 1969 1 and, since then, has undergone many modifications. The most commonly used technique was described by Seglenin 1976 2. Essentially, hepatocytes are dissociated from anesthetized adult rats by a non-recirculating collagenase perfusion through the portal vein. The isolated cells are then filtered through a 100 μm pore size mesh nylon filter, and cultured onto plates. After 4-hour culture, the medium is replaced with serum-containing or serum-free medium, e.g. Hepato ZYME-SFM, for additio nal time to culture. These procedures require surgical and sterile culture steps that can be better demonstrated by video than by text. Here, we document the detailed steps for these procedures by both video and written protocol, which allow consistently in the generation of viable hepatocytes in large numbers.

Original languageEnglish (US)
Article numbere3917
JournalJournal of Visualized Experiments
Issue number64
StatePublished - Jun 29 2012
Externally publishedYes


  • Cellular biology
  • Hepatic cells
  • Hepatocyte
  • Issue 64
  • Medicine
  • Physiology
  • Primary cell culture
  • Rat

ASJC Scopus subject areas

  • General Neuroscience
  • General Chemical Engineering
  • General Biochemistry, Genetics and Molecular Biology
  • General Immunology and Microbiology


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