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J Biosci Bioeng. 2006 Jan;101(1):26-30.

Effect of oxygen on in vitro differentiation of mouse embryonic stem cells.

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Division of Medicine and Engineering Science, Interdisciplinary Graduate School of Medicine and Engineering, University of Yamanashi, 4-3-11 Takeda, Kofu, Yamanashi 400-8511, Japan.


The monolayer culture of embryonic stem (ES) cells and embryoid body (EB) formation were carried out under various oxygen tensions (i.e., 5% O2, 20% O2, and 40% O2). The ES cells cultured in 5% O2 and 20% O2 exhibited decreased specific alkaline phosphatase (AP) activity with culture time, whereas the ES cells cultured in 40% O2 retained their specific AP activity until culture day 3. The ES cells in 40% O2 maintained an octamer-binding transcription factor 4 (Oct-4; a marker of pluripotent undifferentiated ES cells) gene expression level higher than that at other oxygen tensions. The EB formed in 40% O2 maintained an Oct-4 gene expression level higher than that at other oxygen tensions. The generation of cardiomyocytes and the decline in Oct-4 gene expression level were earliest in the EB formed in 20% O2. It was suggested that the culture condition under a high oxygen tension (40% O2) retards the differentiation of ES cells and a normal oxygen tension (20% O2) allows spontaneous cell differentiation.

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