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Effects of Oxygen Concentration on the Proliferation and Differentiation of Mouse Neural Stem Cells In Vitro

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Abstract

Background and purpose Cerebral ischemia is known to elicit the activation of neural stem cells (NSCs); however its mechanism is not fully determined. Although oxygen concentration is known to mediate many ischemic actions, there has been little attention given to the role of pathological oxygen changes under cerebral ischemia on the activation of NSCs. We investigated the effects of various oxygen concentrations on mouse neural stem cells in vitro. Methods NSCs were cultured from the ganglionic eminence of fetal ICR mice on embryonic day 15.5 using a neurosphere method. The effects of oxygen concentrations on proliferation, differentiation, and cell death of NSCs were evaluated by bromodeoxyuridine (BrdU) incorporation, immunocytochemistry, and TUNEL assay, respectively. Results The highest proliferation and the neuronal differentiation of the NSCs were observed in 2% oxygen, which yielded significantly higher proportions of both BrdU-labeled cells and Tuj1-positive cells when compared with 20% and 4% oxygen. On the other hand, the differentiation to the astrocytes was not affected by oxygen concentrations, except in the case of anoxia (0% oxygen). The cell death of the NSCs increased in lower oxygen conditions and peaked at anoxia. Furthermore, the switching of the neuronal subtype differentiation from GABA-positive to glutamate-positive neurons was observed in lower oxygen conditions. Conclusions These findings raise the possibility that reduced oxygen levels occurring with cerebral ischemia enhance NSC proliferation and neural differentiation, and that mild hypoxia (2% oxygen), which is known to occur in the ischemic penumbra, is suitable for abundant neuronal differentiation.

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Acknowledgments

The authors thank Anne-Lise Maag and William Slikker (Department of Neurosurgery, Stanford University) for editing the manuscript.

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Authors and Affiliations

  1. Department of Neurobiology and Behavior, Unit of Basic Medical Sciences, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki, 852-8523, Japan

    Nobutaka Horie, Kenji So, Takahiro Moriya & Kazuyuki Shinohara

  2. Department of Neurosurgery, Nagasaki University Graduate School of Medicine, Nagasaki, Japan

    Nobutaka Horie, Naoki Kitagawa, Keisuke Tsutsumi & Izumi Nagata

Authors
  1. Nobutaka Horie
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  2. Kenji So
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  3. Takahiro Moriya
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  4. Naoki Kitagawa
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  5. Keisuke Tsutsumi
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  6. Izumi Nagata
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  7. Kazuyuki Shinohara
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Correspondence to Nobutaka Horie or Kazuyuki Shinohara.

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Horie, N., So, K., Moriya, T. et al. Effects of Oxygen Concentration on the Proliferation and Differentiation of Mouse Neural Stem Cells In Vitro. Cell Mol Neurobiol 28, 833–845 (2008). https://doi.org/10.1007/s10571-007-9237-y

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  • DOI: https://doi.org/10.1007/s10571-007-9237-y

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