Currently at Department of Biology, Temple University, Philadelphia.
Effects of Ethanol on Mouse Embryonic Stem Cells
Article first published online: 17 SEP 2009
Copyright © 2009 by the Research Society on Alcoholism
Alcoholism: Clinical and Experimental Research
Volume 33, Issue 12, pages 2172–2179, December 2009
How to Cite
Arzumanyan, A., Anni, H., Rubin, R. and Rubin, E. (2009), Effects of Ethanol on Mouse Embryonic Stem Cells. Alcoholism: Clinical and Experimental Research, 33: 2172–2179. doi: 10.1111/j.1530-0277.2009.01057.x
- Issue published online: 19 NOV 2009
- Article first published online: 17 SEP 2009
- Received for publication March 24, 2009; accepted July 31, 2009.
Vol. 35, Issue 4, 773, Article first published online: 10 NOV 2010
- Stem Cells;
- Transcription Factors
Background: Fetal alcohol syndrome (FAS) reflects a constellation of congenital abnormalities caused by excess maternal consumption of alcohol. It is likely that interference with embryonic development plays a role in the pathogenesis of the disorder. Ethanol-induced apoptosis has been suggested as a causal factor in the genesis of FAS. Mouse embryonic stem (mES) cells are pluripotent cells that differentiate in vitro to cell aggregates termed embryoid bodies (EBs), wherein differentiation capacity and gene expression profile are similar to those of the early embryo.
Methods: To investigate the effects of ethanol during differentiation, mES cells were cultured on a gelatin surface in the presence of leukemia inhibitory factor which maintains adherent undifferentiated cells or in suspension to promote formation of EBs. All cells were treated (1–6 days) with 80 mM ethanol. The pluripotency and differentiation of mES cells were evaluated by western blotting of stage-specific embryonic antigen (SSEA-1), transcription factors Oct-3/4, Sox-2, and Nanog, using alkaline phosphatase staining. Apoptosis (early to late stages) was assessed by fluorescence-activated cell sorting using TdT-mediated biotin–dUTP nick-end labelling assay and fluorescein isothiocyanate-Annexin V/propidium iodide staining.
Results: Ethanol increased apoptosis during in vitro differentiation of mES cells to EBs, whereas undifferentiated cells were not affected. Ethanol exposure also interfered with pluripotency marker patterns causing an upregulation of SSEA-1 under self-renewal conditions. In EBs, ethanol delayed the downregulation of SSEA-1 and affected the regulation of transcription factors during differentiation.
Conclusion: Our findings suggest that ethanol may contribute to the pathogenesis of FAS by triggering apoptotic pathways during differentiation of embryonic stem cells and deregulating early stages of embryogenesis.