Time-Related Analysis of Metabolic Liver Functions, Cellular Morphology, and Gene Expression of Hepatocytes Cultured in the Bioartificial Liver of the Academic Medical Center in Amsterdam (AMC-BAL)

A comprehensive understanding of the mechanisms that underlie hepatic differentiation inside a bioartificial liver (BAL) device is obtained when functional, histological, and gene expression analyses can be combined. We therefore developed a novel cell-sampling technique that enabled us to analyze a...

Full description

Saved in:
Bibliographic Details
Published in:Tissue engineering 2007-06, Vol.13 (6), p.1235-1246
Main Authors: Poyck, Paul P.C., Hoekstra, Ruurdtje, Chhatta, Aniska, Bloemendaal, Lysbeth Ten, van Wijk, Albert C.W.A., Galavotti, Daniele, van Gulik, Thomas M., Chamuleau, Robert A.F.M.
Format: Article
Language:English
Subjects:
Academic Medical Centers
Animals
Biomedical materials
Cell culture
Cell Culture Techniques - instrumentation
Cell Culture Techniques - methods
Cell Proliferation
Cell Size
Cells, Cultured
Cellular biology
Equipment Design
Equipment Failure Analysis
Gene expression
Gene Expression Regulation - physiology
Hepatocytes - cytology
Hepatocytes - metabolism
Liver
Liver - cytology
Liver - metabolism
Liver, Artificial
Netherlands
Proteome - metabolism
Swine
Tissue engineering
Tissue Engineering - instrumentation
Tissue Engineering - methods
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:A comprehensive understanding of the mechanisms that underlie hepatic differentiation inside a bioartificial liver (BAL) device is obtained when functional, histological, and gene expression analyses can be combined. We therefore developed a novel cell-sampling technique that enabled us to analyze adherent hepatocytes inside a BAL device during a 5-day culture period, without the necessity of terminating the culture. Biochemical data showed that hepatocyte-specific functions were relatively stable, despite an increase in glycolytic activity. Quantitative reverse transcriptase polymerase chain reaction analysis of hepatic genes cytochrome p450 3A29, albumin, glutamine synthetase, alpha-1 antitrypsin, and carbamoyl-phosphate synthetase, but also de-differentiation marker π-class glutathione S transferase showed stable messenger ribonucleic acid (mRNA) levels from day 1 to 5. In contrast, mRNA levels of α-fetoprotein, pro- and anti-apoptotic genes Bax-α and Bcl-X L , metabolic genes lactate dehydrogenase and uncoupling protein 2, and cytoskeleton genes α- and β-tubulin and β-actin increased in 5 days. Histological analysis revealed viable tissue-like structures with adaptation to the in vitro environment. We conclude that hepatocytes show a tendency for de-differentiation shortly after seeding but thereafter remain acceptably differentiated during 5 days of culture. Furthermore, partly impaired mitochondrial function is suggestive for local hypoxic regions and may trigger the observed metabolic changes. Anti-apoptotic activity seems to balance pro-apoptotic activity. This new cell-sampling technique facilitates the analysis of dynamic processes of hepatocyte culture inside a BAL.
ISSN:1076-3279
1557-8690
DOI:10.1089/ten.2006.0343