Rapid isolation of murine primary hepatocytes for chromosomal analysis

Kentaro Ariyoshi, Yohei Fujishima, Tomisato Miura, Yi Shang, Shizuko Kakinuma, Shimada Yoshiya, Kosuke Kasai, Akifumi Nakata, Akira Tachibana, Mitsuaki A. Yoshida

Research output: Contribution to journalArticlepeer-review

2 Citations (Scopus)

Abstract

Primary hepatocyte culture is a crucial tool for investigations of liver function and for evaluating the toxic effects of drugs. In addition, chromosomal analysis of hepatocytes could also prove useful for understanding the mechanisms of hepatocarcinogenesis. However, cultivation of primary hepatocytes for chromosome analysis has been hampered by the specific equipment and skill required to perform the in situ perfusion step necessary for isolation of primary hepatocytes. In the present study, we aimed to establish a simple and efficient method of isolating hepatocytes suitable for chromosome analysis. We performed hepatocyte isolation without using collagenase perfusion, instead digesting liver tissues using collagenase in tubes. In addition, we examined hepatocyte and bone marrow cell (BMC) co-culture and cultivation of hepatocytes with medium containing BMC culture medium supernatants. We found that hepatocyte viability and attachment rate were significantly improved, both by co-culture with BMCs and medium containing BMC culture media supernatants, with the latter also significantly increasing the mitotic index. Using this simple method of isolation and cultivation, we could successfully perform chromosomal analysis of mouse primary hepatocytes. This method has the potential to help understand the mechanisms underlying chromosomal instability-mediated hepatocarcinogenesis.

Original languageEnglish
Pages (from-to)474-478
Number of pages5
JournalIn Vitro Cellular and Developmental Biology - Animal
Volume53
Issue number5
DOIs
Publication statusPublished - 2017 May 1
Externally publishedYes

Keywords

  • Chromosome analysis
  • Co-culture
  • Hepatocyte

ASJC Scopus subject areas

  • Developmental Biology
  • Cell Biology

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