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Cytotechnology

, Volume 33, Issue 1–3, pp 53–61 | Cite as

Establishment and characterization of immortalized hippocampal neural precursor cell lines

  • Tomohiro Kaji
  • Naoko Yoshida
  • Kiyoshi Yamada
  • Tatsuhiro Hisatsune
  • Shuichi Kaminogawa
Article

Abstract

In the mammalian central nervous system, a complexcircuit of neurons contributes to higher behaviors.Each region of the brain has a unique function derivedfrom various types of neurons. Several neuralprecursor cell lines have been established from basalganglia of fetal brain. In this study, hippocampalneural precursor cell lines were established from thehippocampus of p53-/- embryos. By means ofintegration of a MycER regulatable oncoprotein intop53-/- neural precursor cells, several immortallines were established from embryonic hippocampalprimordium, with bFGF and estrogen continuouslysupplied for activation of the MycER protein. A dualluciferase study demonstrated that the MycER proteinblocked the expression of a glial cell marker protein,GFAP, probably contributing to the persistent celldivision of the immortalized neural precursor cells.These cell lines differentiate into neuronal and glialcell types after withdrawal of bFGF. The phenotype ofthe hippocampal cell lines differed from that of thebasal ganglia cell lines as observed in a clonaldensity culture. This result implies that each regionof the brain has a unique developmental program, thatmay be imprinted in each of the neural precursor cells.

brain development brain repair hippocampus immortalization Myc neural precursor cells p53 

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References

  1. Blackwell TK, Kreztner L, Blackwood EM, Eisenman RN and Weintraub H (1990) Sequence-specific DNA binding by the c-MYC protein. Science 250: 1149–1151.Google Scholar
  2. Cattaneo E and McKay RDG (1990) Proliferation and differentiation of neuronal stem cells regulated by nerve growth factor. Nature 347: 762–765.Google Scholar
  3. Caviness Jr VS, Takahashi T and Nowakowski RS (1995) Numbers, time and neocortical neuronogenesis: A general developmental and evolutionary model. Trends Neurosci 18: 379–383.Google Scholar
  4. Eilers M, Picard D, Yamamoto K and Bishop JM (1989) Chimaeras of Myc oncoprotein and steroid receptors cause hormonedependent transformation of cells. Nature 340: 66–68.Google Scholar
  5. Finlay BL and Darlington RB (1995) Linked regularities in the development and evolution of mammalian brain. Science 268: 1578–1584.Google Scholar
  6. Frederiksen K, Jat PS, Valtz N, Levy D and McKay RDG (1988) Immortalization of precursor cells from the mammalian CNS. Neuron 1: 439–448.Google Scholar
  7. Hoshimaru M, Ray J, Sah DWY and Gage FH (1996) Differentiation of the immortalized adult neuronal progenitor cell line HC2S2 into neurons by regulatable suppression of the v-myc oncogene. Proc Natl Acad Sci USA 93: 1518–1523.Google Scholar
  8. Jacobson M (1991) Developmental Neurobiology. Plenum Press, New York.Google Scholar
  9. Jansen-Durr P, Meichle A, Steiner P, Pagano M, Finke K, Botz J, Wessbecher J, Draetta G and Eilers M (1993) Differential modulation of cyclin gene expression by MYC. Proc Natl Acad Sci USA 90: 3685–3689.Google Scholar
  10. Johe KK, Hazel TG, Muller T, Dugich-Djordjevic MM and McKay RDG (1996) Single factors direct the differentiation of stem cells from the fetal and adult central nervous system. Genes Dev 10: 3129–3140.Google Scholar
  11. Levine AJ (1997) p53, the cellular gatekeeper for growth and division. Cell 88: 323–331.Google Scholar
  12. Lundberg C, Martinez-Serrano A, Cattaneo E, McKay RDG and Bjorklund A (1997) Survival, integration, and differentiation of neural stem cell lines after transplantation to adult rat striatum. Exp Neurol 145: 342–360.Google Scholar
  13. Martinez-Serrano A and Bjorklund A (1997) Immortalized neural progenitor cells for CNS gene transfer and repair. Trends Neurosci 20: 530–538.Google Scholar
  14. McKay RDG (1997) Stem cells in the central nervous system. Science 276: 66–71.Google Scholar
  15. Metz T, Harris AW and Adams JM (1995) Absence of p53 allows direct immortalization of hematopoietic cells by the myc and raf oncogenes. Cell 82: 29–36.Google Scholar
  16. Miura M, Tamura T and Mikoshiba K (1990) Cell-specific expression of the mouse glial fibrillary acidic protein gene: identification of the cis-and trans-acting promoter elements for astrocyte-specific expression. J Neurochem 55: 1180–1188.Google Scholar
  17. Mizushima S and Nagata S (1990) pEF-BOS, a powerful mammalian expression vector. Nucleic Acids Res 18: 5322.Google Scholar
  18. Monaghan AP, Bock D, Gass P, Schwager A, Wolfer DP, Lipp H-P and Schutz G (1997) Defective limbic system in mice lacking the tailless gene. Nature 390: 515–517.Google Scholar
  19. Nakafuku M and Nakamura S (1995) Establishment and characterization of a multipotential neural cell line that can conditionally generate neurons, astrocytes, and oligodendrocytes in vitro. J Neurosci Res 41: 153–168.Google Scholar
  20. Snyder EY, Deitcher DL, Walsh C, Arnold-Aldea S, Hartwieg EA and Cepko CL (1992) Multipotent neural cell lines can engraft and participate in development of mouse cerebellum. Cell 68: 33–51.Google Scholar
  21. Suhonen JO, Peterson DA, Ray J and Gage FH (1996) Differentiation of adult hippocampus-derived progenitors into olfactory neurons in vivo. Nature 383: 624–627.Google Scholar
  22. Timme TL and Thompson TC (1994) Rapid allelotype analysis of p53 knockout mice. BioTechniques 17: 461–463.Google Scholar
  23. Vicario-Abejon C, Johe KK, Hazel TG, Collazo D and McKay RDG (1995) Functions of basic fibroblast growth factor and neurotrophins in the differentiation of hippocampal neurons. Neuron 15: 105–114.Google Scholar
  24. Yamada K, Hisatsune T, Uchino S, Nakamura T, Kudo Y and Kaminogawa S (1999) NMDA receptor-mediated Ca2+ responses in neurons differentiated from p53—/— immortalized neural stem cells. Neurosci Lett 264: 165–167.Google Scholar

Copyright information

© Kluwer Academic Publishers 2000

Authors and Affiliations

  • Tomohiro Kaji
    • 1
  • Naoko Yoshida
    • 1
  • Kiyoshi Yamada
    • 1
  • Tatsuhiro Hisatsune
    • 1
  • Shuichi Kaminogawa
    • 1
  1. 1.Department of Applied Biological ChemistryThe University of TokyoTokyoJapan

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