Expression of α2-macroglobulin by the interaction between hepatocytes and endothelial cells in coculture

Mark A. Talamini, Michael P. McCluskey, Timothy G. Buchman, Antonio De Maio

Research output: Contribution to journalArticle

Abstract

The interaction between distinct cell types within the liver seems to be important in regulating hepatic function. However, these interactions have not been well characterized because of difficulty in reproducing the hepatic environment in an ex vivo model. In the present study a coculture system of hepatocytes and endothelial cells was established to investigate the communication between parenchymal and nonparenchymal cells. Freshly isolated rat hepatocytes were placed onto a monolayer of primary aortic rat endothelial cells. Analysis of the proteins secreted into the extracellular medium after pulse labeling with radioactive amino acids revealed the presence of a 180,000-apparent molecular weight glycoprotein, BBB-180, which was not detected in the extracellular medium of hepatocytes or endothelial cells when they were cultured separately. This glycoprotein was identified as α2-macroglobulin after sequencing of the proteolytic peptides derived from the purified protein. This finding was confirmed by Northern and Western blotting, immunoprecipitation, and RT-PCR. The expression of α2- macroglobulin required direct contact between hepatocytes and viable endothelial cells. These findings suggest that endothelial cells modulate hepatocyte gene expression by direct cellular interactions.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Regulatory Integrative and Comparative Physiology
Volume275
Issue number1 44-1
StatePublished - Jul 1998

Fingerprint

Macroglobulins
Coculture Techniques
Hepatocytes
Endothelial Cells
Liver
Glycoproteins
Immunoprecipitation
Northern Blotting
Proteins
Molecular Weight
Western Blotting
Communication
Gene Expression
Amino Acids
Polymerase Chain Reaction
Peptides

Keywords

  • Acute phase
  • Cellular communication
  • Liver

ASJC Scopus subject areas

  • Physiology
  • Physiology (medical)

Cite this

Expression of α2-macroglobulin by the interaction between hepatocytes and endothelial cells in coculture. / Talamini, Mark A.; McCluskey, Michael P.; Buchman, Timothy G.; De Maio, Antonio.

In: American Journal of Physiology - Regulatory Integrative and Comparative Physiology, Vol. 275, No. 1 44-1, 07.1998.

Research output: Contribution to journalArticle

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