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"The roles of neurotrophic factors and nitric oxide on the differentiation of human neuronal cells"

Research Project

Project/Area Number 08671584
Research Category

Grant-in-Aid for Scientific Research (C)

Allocation TypeSingle-year Grants
Section一般
Research Field Cerebral neurosurgery
Research InstitutionKobe University

Principal Investigator

KONDOH Takeshi  Department of Neurosurgery Kobe University School of Medicine, 医学部, 助手 (50273769)

Co-Investigator(Kenkyū-buntansha) TAMAKI Norihiko  Department of Neurosurgery Kobe University School of Medicine, 医学部, 教授 (10030941)
Project Period (FY) 1996 – 1997
Project Status Completed (Fiscal Year 1997)
Budget Amount *help
¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 1997: ¥900,000 (Direct Cost: ¥900,000)
KeywordsNeuronal Stem Cell / Teratocarcinoma / Retinoic Acid / Transplantation / Astrocytes / GABA receptor / Apoptosis / Blood-Brain-Barrier / Cryopreservation / retinoic acid / whole-cell patch-clamp / アセチルコリン受容体 / 神経細胞移植 / NADPH-d / NT-2
Research Abstract

In the present study, we have first examined the possibility of neuronal differentiation of human teratocartinoma cell line (NT-2) with a purpose of alternative source for neuronal transplantation. By means of treating the teratocarcinoma cells (NT-2) in vitro with 10 mM of retinoic acid for five weeks, neuronal like cells in morphology (NT-2N) were successfully obtained. NT-2N cells as well as NT-2 cells were then studied electrophysiologicaly for their property as neuronal cells. We have found that voltage sensitive Na+ currents were observed in NT-2N cells but not in the NT-2 cells and that in addition to the glutamate receptors, the NT-2N cells expressed GABA-A receptors, suggesting that those receptors have a crucial role in neurotansmission from the earlier stage of the brain development.
Second, neural stem cells isolated from rat fetus striatum were cultured in serum free epidermal growth factor containing medium as spheres for two months without dissociation. Each nonpassaged s … More phere maintained the potential to differentiate to neurons and glia cells when replated in serum containing medium. The electronmicroscopical study revealed immature cells with scant dark cytoplasm or in proliferation stage. The cells differentiated heterogeneously in the sphere ; some had rough endoplasmic reticula and others had long processes with intermediate filaments. Cells undergoing apoptosis were also observed in the sphere. Those were dominant in the core of spheres where abundant cellular debris including apoptotic bodies were observed in the large extracellular spaces. These neural spheres with ongoing differentiation of the cells could fit as an alternative source for neuonal transplantation.
Third, a human astrocyte cell line was generated from a tissue of white matter containing reactive astrocytes in a routine neurosurgical procedure. Those astrocytes were cryopreserved and grafted into normal rat striatum with immunosurpression by cycrosporine. Immunohistochemical study with human specific glial fibrillary acidic protein antigen revealed that grafted astrocytes formed dense cell clusters in the host brain and that part of the cells migrated away into the host striatum. Inside the grafts, there were vessels which were positive with rat specific endothelial barrier antigen (EBA), suggesting that the angiogenesis of the grafts was performed by the host-derived endothelial cells. The expression of EBA also suggests that the blood-brain-barrier was likely formed in the xenograft of astrocytes. In the animals in which the immunosurpression discontinued at two weeks postgrafting, the grafts were able to survive for three additional weeks, however, rejected at eight weeks after discontinuation of the immunosurpression. This indicated that the barrier formation as an immunological privilege was incomplete. Our study indicates that although the formation of blood-brain-barrier is incomplete in the xenografts of astrocytes, the grafted astrocytes themselves were remarkablely integrated with host circumstances and that the autograft of the astrocytes might be feasible for the therapeutic purpose. Less

Report

(3 results)
  • 1997 Annual Research Report   Final Research Report Summary
  • 1996 Annual Research Report
  • Research Products

    (19 results)

All Other

All Publications (19 results)

  • [Publications] Matsuoka T.et al.: "The GABA_A receptor is expressed in human neurons derived from a feratocarcinoma cell line" Biochemical and Biophysical Research Communication. 237. 719-723 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 近藤 威: "神経・筋疾患に対する細胞移植" 脳と神経. 49. 507-515 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 近藤 威: "長期培養神経幹細胞の増殖能および分化能に関する研究" 神経組織の成長・再生・移植. 9. 97-98 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 秋山 英之: "ヒトアストロサイトのラット脳内移植后の組織像" 神経組織の成長・再生・移植. 9. 61-62 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] 西崎 知之: "Teracocarcinoma cell(NT2)より分化させたヒト神経細胞におけるGABA_A受容体の発現" 神経組織の成長・再生・移植. 9. 71-72 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Kondoh, T.et al.: "Expression of Neurotransmitter receptor and grafting of human neuron derived from NT2 cells" Soc.Neurosci.Abstr.23. 345 (1997)

    • Description
      「研究成果報告書概要(和文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Matsuoka Toshiyuki, Takeshi Kondoh, Norihiko Tamaki, Tomoyuki Nishizaki: "The GABA-A receptor is expressed in human neurons derived from a teratocarcinoma cell line." Biochemical and Biophysical Research Communication. 237. 719-723 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Kondoh, T.Nishizaki, N.Tamaki, T.Matsuoka, H.Akiyama, M.Mori, Y.Okada: "Expression of neurotransmitter receptor and grafting of human neurons derived from NT-2 cells." Soc Neurosci Abstr. 23 Part1. 345 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] Takeshi Kondoh, Norihiko Tamaki: "Cell transplantation for neuronal and muscular disorders." Brain and Nerve. 49. 507-515 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] N.Nishizaki, T Kondoh, T.Matsuoka, N.Tamaki: "GABA-A receptor is expressed in human neurons derived from a teratocarcinoma cell line (NT2)." The Abstracts for the Japanese Society for Neural Growth, Regeneration and Transplantation. 9. 71-72 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] H.Akiyama, T.Kondoh, T.Kokunai, N.Tamaki: "Immunohistochemical study of human astrocytes grafted into rat striatum" The Abstracts for the Japanese Society for Neural Growth, Regeneration and Transplantation. 9. 61-62 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] T.Kondoh, E.Fujimoto, H.Akiyama, N.Tamaki: "Prolireration and differentiation of long-term cultured nonpassage neural spheres" The Abstracts for the Japanese Society for Neural Growth, Regeneration and Transplantation. 9. 97-98 (1997)

    • Description
      「研究成果報告書概要(欧文)」より
    • Related Report
      1997 Final Research Report Summary
  • [Publications] MATUSOKA T. et al,: "The GABA_A receptor is expressed in human neurons derived from a feratocacinoma cell line." Biochemical and Biophysical Research Communication. 237. 719-723 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 近藤威: "神経・筋疾患に対する細胞移植" 脳と神経. 49. 507-515 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 近藤威: "長期培養神経幹細胞の増殖能および分化能に関する研究" 神経組織の成長・再生・移植. 9. 97-98 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 秋山英之: "ヒトアストロサイトのラット脳内移植后の組織像" 神経組織の成長・再生・移植. 9. 61-62 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 西崎知之: "Teratocarcinome cell line(NT2)より分化させたヒト神経細胞におけるGABA_A受容体の発現" 神経組織の成長・再生・移植. 9. 71-72 (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] Kondoh,T.et al: "Expression of neurotransmitter ceptor and grafting of human neurons derived from NT2 cells" Soc.Neurosci.Abstr.23. 345- (1997)

    • Related Report
      1997 Annual Research Report
  • [Publications] 近藤 威、玉木紀彦: "神経・筋疾患に対する細胞移植" 脳と神経. (印刷中). (1997)

    • Related Report
      1996 Annual Research Report

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Published: 1997-04-01   Modified: 2016-04-21  

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