Novel method to examine hepatocyte-specific gene expression in a functional coculture system

Yasunori Kurosawa, Akiyoshi Taniguchi*, Teruo Okano

*この研究の対応する著者

研究成果: Article査読

20 被引用数 (Scopus)

抄録

To mimic native tissue function, cocultare systems are an extremely useful model. In many cases, differentiated functions can be maintained only through the interactions of various cell types. Therefore, methods for examining the interactions between cocultured cells are necessary. The assessment of cell-to-cell cross-talk at the level of gene expression is one such method to examine interactions between different cell types. However, it is generally difficult to determine the gene expression of specific cell types in coculture without first separating cell populations. To overcome these obstacles, we have established a novel method to determine gene expression levels of a targeted cell population in coculture, using species-specific primers. With this approach, we were able to determine hepatocyte-specific gene expression of Fao cells (a rat hepatocyte cell line) in culture with human umbilical vein endothelial cells (HUVECs). Expression of both albumin and apolipoprotein A-I (apoA-I) increased time dependently for 10 days and maintained significantly higher expression in the coculture system as compared with isolated Fao cells. This indicates that hepatocyte function increased gradually in our coculture system and could be maintained long-term, suggesting that the construction of mature cell-to-cell communication between the two cell lines required a considerable amount of time. The expression of HNF-4 and HNF-1α, which are liver-enriched transcription factors, did not differ between the monolayer and cocultured Fao cells, suggesting that expression of HNF-4 and HNF-1α was not responsible for the increased expression albumin and apoA-I. Our findings suggest that this novel method for the detection of gene expression of targeted cell populations can be a useful tool in determining the molecular mechanisms that regulate communication between different cell types.

本文言語English
ページ(範囲)1650-1657
ページ数8
ジャーナルTissue Engineering
11
11-12
DOI
出版ステータスPublished - 2005 11月 1
外部発表はい

ASJC Scopus subject areas

  • バイオテクノロジー
  • 生物理学
  • 細胞生物学

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