Document Detail

Lowering oxygen tension enhances the differentiation of mouse embryonic stem cells into neuronal cells.
MedLine Citation:
PMID:  19455654     Owner:  NLM     Status:  MEDLINE    
Embryonic stem cells (ESC) are capable of proliferating indefinitely in vitro whilst retaining their ability to differentiate into cells of every adult lineage. Efficient, high yield processes, which direct differentiation of ESC to specific lineages, will underpin the development of cost-effective drug screening and cell therapy products. The aim of this study was to investigate whether laboratory oxygen tension currently used for the neuronal differentiation of ESC was suboptimal resulting in inefficient process yields. An adherent monolayer protocol for the neuronal differentiation of mouse ESC (mESC) was performed in oxygen controlled chambers using a chemically defined media over an 8 day period of culture. When exposed to oxygen tensions more appropriate to in vivo neuronal development (2% O(2)), there was a 34-fold increase in the yield of viable cells from the differentiation process. Low oxygen tension inhibited cell death during an early phase (48 to 96 h) and toward the end (120 to 192 h) of the process. The percentage of cells expressing neuronal markers was determined by flow cytometry, revealing a small rise in the betaIII tubulin and a threefold increase in the MAP2 populations at 2% O(2). The total increase in the yield of viable cells expressing neuronal markers was shown to be 55-fold for betaIII tubulin and 114-fold for MAP2. In conclusion, this study revealed that low oxygen tension can be used to enhance the yield of neuronal cells derived from ESCs and has implications for the development of efficient, cost-effective production processes.
Paul Mondragon-Teran; Gary J Lye; Farlan S Veraitch
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology progress     Volume:  25     ISSN:  1520-6033     ISO Abbreviation:  Biotechnol. Prog.     Publication Date:    2009 Sep-Oct
Date Detail:
Created Date:  2009-11-05     Completed Date:  2010-02-22     Revised Date:  2011-06-29    
Medline Journal Info:
Nlm Unique ID:  8506292     Medline TA:  Biotechnol Prog     Country:  United States    
Other Details:
Languages:  eng     Pagination:  1480-8     Citation Subset:  IM    
Copyright Information:
2009 American Institute of Chemical Engineers Biotechnol.
The Advanced Centre for Biochemical Engineering, Dept. of Biochemical Engineering, University College London, UK.
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MeSH Terms
Cell Culture Techniques / methods
Cell Survival
Embryonic Stem Cells / cytology,  metabolism,  physiology*
Flow Cytometry
Intermediate Filament Proteins / metabolism
Microscopy, Phase-Contrast
Microtubule-Associated Proteins / metabolism
Nerve Tissue Proteins / metabolism
Neurogenesis / physiology*
Neurons / cytology,  metabolism*
Oxygen / chemistry*
Rosette Formation
Tubulin / metabolism
Reg. No./Substance:
0/Intermediate Filament Proteins; 0/Microtubule-Associated Proteins; 0/Mtap2 protein, mouse; 0/Nerve Tissue Proteins; 0/Tubulin; 0/nestin; 7782-44-7/Oxygen

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