Abstract |
The instability of the hepatocyte phenotype in vitro has limited the ability to quantitatively investigate regulation of stress responses of the liver. Here, we adopt a tissue-engineering approach to form stable liver tissue in vitro by forming collagen "sandwich" cultures of transgenic murine hepatocytes harboring a regulatory gene of interest flanked by loxP sites. The floxed gene is excised in a subset of cultures by transfection with adenovirus carrying the gene for Cre-recombinase, thereby generating wild-type and null liver tissues from a single animal. In this study, we specifically investigated the role of hypoxia inducible factor 1 alpha (HIF-1alpha) in the hepatocellular response to hypoxia. Using high-density oligonucleotide arrays, we examined genome-wide gene expression after 8 h of hypoxia in wild-type and HIF- 1alpha null hepatocyte cultures. We identified more than 130 genes differentially expressed under hypoxia involved in metabolic adaptation, angiogenic signaling, immediate early response, and cell cycle regulation. Real-time polymerase chain reaction analysis verified that known hypoxia-responsive genes such as glucose transporter-1 and vascular endothelial growth factor were induced in a HIF-1alpha-dependent manner under hypoxia. Our results demonstrate the potential to integrate in vitro tissue models with transgenic and microarray technologies for the study of physiologic stress responses.
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Authors | Jared W Allen, Salman R Khetani, Randall S Johnson, Sangeeta N Bhatia |
Journal | Tissue engineering
(Tissue Eng)
Vol. 12
Issue 11
Pg. 3135-47
(Nov 2006)
ISSN: 1076-3279 [Print] United States |
PMID | 17518628
(Publication Type: Journal Article)
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Chemical References |
- Glucose Transporter Type 1
- Hif1a protein, mouse
- Hypoxia-Inducible Factor 1, alpha Subunit
- Vascular Endothelial Growth Factor A
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Topics |
- Adenoviridae
(genetics)
- Animals
- Cell Culture Techniques
- Cell Hypoxia
(physiology)
- Cells, Cultured
- Gene Expression Profiling
- Gene Expression Regulation
- Genome
- Glucose Transporter Type 1
(genetics)
- Hepatocytes
(cytology)
- Hypoxia-Inducible Factor 1, alpha Subunit
(genetics, physiology)
- Liver
(cytology)
- Mice
- Mice, Transgenic
- Models, Biological
- Oligonucleotide Array Sequence Analysis
- Polymerase Chain Reaction
- Tissue Engineering
(methods)
- Vascular Endothelial Growth Factor A
(genetics)
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