An optimal culture condition maintains human hepatocyte phenotype after long-term culture

Naoki Yamamoto, Jian Wu, Yanhong Zhang, Andreea M. Catana, Hongbo Cai, Stephen Strom, Phyllis M. Novikoff, Mark A Zern

Research output: Contribution to journalArticlepeer-review

17 Scopus citations

Abstract

Background: Long-term culture of primary hepatocytes from various species is impeded by a decrease of cell viability and a loss of hepatocyte-specific function. The aim of the present study was to investigate whether our optimal culture condition (OC) can maintain the phenotype of primary hepatocytes in long-term culture. Methods: Primary human hepatocytes were cultured in either hepatocyte maintenance medium (HM) or OC for 2-4 weeks. Expression of hepatocyte-specific genes was determined by real-time quantitative RT-PCR. Results: The level of albumin mRNA in human hepatocytes cultured in OC was 11-fold more than in HM and gene expression levels of α1-antitrypsin and transferrin were at approximately 40 and 11% of freshly isolated primary human hepatocytes. Electron microscopy revealed that cells in OC displayed hepatocyte properties (e.g. polarity, junctional complexes, bile canaliculi and glycogen particles). Cytochrome P4501A1/2 activity of hepatocytes cultured in OC was 15- and 17-fold higher than in HM at 2 and 4 weeks of culture, and DNA synthesis was higher. Conclusions: Using our optimal culture condition, we were able to maintain the phenotype of primary human hepatocytes in long-term culture. They not only maintain better liver-specific function, but also retain higher proliferative potential.

Original languageEnglish (US)
Pages (from-to)169-177
Number of pages9
JournalHepatology Research
Volume35
Issue number3
DOIs
StatePublished - Jul 2006

Keywords

  • Cytochrome P450
  • Hepatocyte
  • Long-term culture
  • Optimal culture condition

ASJC Scopus subject areas

  • Gastroenterology

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