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| Content Provider | World Health Organization (WHO)-Global Index Medicus |
|---|---|
| Author | Wartenberg, M. Acker, H. Bauer, C. Bichet, S. Marti, H. H. Wenger, R. H. Fandrey, J. Gassmann, M. |
| Description | Author Affiliation: Gassmann M ( Physiologisches Insitut der Universität Zürich-Irchel, Switzerland.); |
| Abstract | Blastocyst-derived pluripotent mouse embryonic stem cells can differentiate in vitro to form so-called embryoid bodies (EBs), which recapitulate several aspects of murine embryogenesis. We used this in vitro model to study oxygen supply and consumption as well as the response to reduced oxygenation during the earliest stages of development. EBs were found to grow equally well when cultured at 20% (normoxia) or 1% (hypoxia) oxygen during the first 5 days of differentiation. Microelectrode measurements of pericellular oxygen tension within 13- to 14-day-old EBs (diameter 510-890 micron) done at 20% oxygen revealed efficient oxygenation of the EBs' core region. Confocal laser scanning microscopy analysis of EBs incubated with fluorescent dyes that specifically stain living cells confirmed that the cells within an EB were viable. To determine the EBs' capability to sense low oxygen tension and to specifically respond to low ambient oxygen by modulating gene expression we quantified aldolase A and vascular endothelial growth factor (VEGF) mRNAs, since expression of these genes is upregulated by hypoxia in a variety of cells. Compared with the normoxic controls, we found increased aldolase A and VEGF mRNA levels after exposing 8- to 9-day-old EBs to 1% oxygen. We propose that EBs represent a powerful tool to study oxygen-regulated gene expression during the early steps of embryogenesis, where the preimplantation conceptus resides in a fluid environment with low oxygen tension until implantation and vascularization allow efficient oxygenation. |
| ISSN | 00278424 |
| e-ISSN | 10916490 |
| Journal | Proceedings of the National Academy of Sciences of the United States of America |
| Issue Number | 7 |
| Volume Number | 93 |
| Language | English |
| Publisher | National Academy of Sciences |
| Publisher Date | 1996-05-01 |
| Publisher Place | United States |
| Access Restriction | Open |
| Subject Keyword | Blastocyst Physiology Cell Differentiation Gene Expression Regulation Oxygen Consumption Stem Cells Aerobiosis Animals Cytology Cell Hypoxia Cells, Cultured DNA Primers Endothelial Growth Factors Biosynthesis Fluorescent Dyes Fructose-Bisphosphate Aldolase Isoenzymes Kinetics Lymphokines Mice Microscopy, Confocal Molecular Sequence Data Polymerase Chain Reaction RNA, Messenger Time Factors Vascular Endothelial Growth Factor A Vascular Endothelial Growth Factors Comparative Study Research Support, Non-U.S. Gov't Multidisciplinary |
| Content Type | Text |
| Resource Type | Article |
| Subject | Multidisciplinary |
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