A bipotent neural progenitor cell line cloned from a cerebellum of an adult p53-deficient mouse generates both neurons and oligodendrocytes

Authors

  • Mitsutoshi Tominaga,

    1. Department of Biological Science and Technology and Tissue Engineering Research Center, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278–8510, Japan
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  • Shinya Honda,

    1. Department of Biological Science and Technology and Tissue Engineering Research Center, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278–8510, Japan
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  • Atsumasa Okada,

    1. Department of Biological Science and Technology and Tissue Engineering Research Center, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278–8510, Japan
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  • Akifumi Ikeda,

    1. Department of Biological Science and Technology and Tissue Engineering Research Center, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278–8510, Japan
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  • Seiji Kinoshita,

    1. Department of Biological Science and Technology and Tissue Engineering Research Center, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278–8510, Japan
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  • Yasuhiro Tomooka

    1. Department of Biological Science and Technology and Tissue Engineering Research Center, Tokyo University of Science, 2641 Yamazaki, Noda, Chiba 278–8510, Japan
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Dr Yasuhiro Tomooka, as above.
E-mail: tomoylab@rs.noda.tus.ac.jp

Abstract

Here we report developmental characteristics of a clonal cell line 2Y-3t established from a multifocal neoplasm that arose in a cerebellum of an adult p53-deficient mouse. The tumorigenicity of the line was not observed in soft agar assay or in nude mouse assay. In serum-containing medium, 2Y-3t cells were epithelial-like in morphology and were mitotic. When they were cultured in serum-free medium, the expressions of neural stem and/or progenitor cell markers were decreased. Concomitantly, the expressions of neuronal and oligodendrocyte markers were increased in concert with morphological differentiation, and DNA synthesis ceased. None of astrocyte markers were detected under these culture conditions. Double-labelling studies revealed that two cell populations coexisted, expressing neuronal or oligodendrocyte markers. Triiodothyronine (T3) increased the oligodendrocyte population when 2Y-3t cells were cultured in serum-free medium. Recloning of the line gave rise to three types of subclones. Sixteen subclones were capable of generating both neurons and oligodendrocytes, four subclones were capable of generating only neurons and one subclone was capable of generating only oligodendrocytes. Thus, 2Y-3t cells have characteristics of bipotent neural progenitor cells capable of generating both neurons and oligodendrocytes. In addition, the line expressed mRNA for Pax-2 and had GAD67-positive cells when cultured in serum-free medium. However, none of the mRNAs for Zic-1, Math1, zebrin or Calbindin-D28k were detected, suggesting that the 2Y-3t line might generate the GABAergic interneuron lineage of the mouse cerebellum.

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