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METHOD FOR PRODUCING NERVE CELLS FROM HUMAN NEURAL STEM CELLS BY DIFFERENTIATION INDUCTION TREATMENT, REAGENT FOR INDUCING DIFFERENTIATION OF NERVE CELLS, AND NERVE CELL DIFFERENTIATION INDUCTION KIT commons meetings

Foreign code F180009476
File No. 2018000986
Posted date Sep 18, 2018
Country WIPO
International application number 2018JP024843
International publication number WO 2019009205
Date of international filing Jun 29, 2018
Date of international publication Jan 10, 2019
Priority data
  • P2017-131308 (Jul 4, 2017) JP
Title METHOD FOR PRODUCING NERVE CELLS FROM HUMAN NEURAL STEM CELLS BY DIFFERENTIATION INDUCTION TREATMENT, REAGENT FOR INDUCING DIFFERENTIATION OF NERVE CELLS, AND NERVE CELL DIFFERENTIATION INDUCTION KIT commons meetings
Abstract The present invention addresses the problem of providing a method for promoting the neural differentiation of neural stem cells. A method for producing nerve cells from human neural stem cells by a differentiation induction treatment, the method including a step of carrying out a differentiation induction treatment of human neural stem cells using a FAD-containing culture medium for nerve cell differentiation induction use.
Outline of related art and contending technology BACKGROUND ART
Mammalian central nervous system neurons and glial cells are present in the produced from neural stem cells. The use of the property of neural stem cells, for the treatment of injured nervous system and realizes a regenerative medicine, drug screening in drug discovery, efficacy and toxicity evaluation of the contribution to the expected.
In recent years, such as a mouse or a rat neural stem cells of rodent, primate including human neural stem cells can be of different nature have been reported (Non-Patent Document 1 and 2). For example, rodent brain cortex of such as neural stem cells located in the band, only to excitatory neurons have differentiated, primate neural stem cells, excitatory neurons inhibitory nerve cell as well as the ability to differentiate into which have been reported.
For application to medical and drug discovery is to, human neural stem cells cultured in a stable, efficient technique for inducing differentiation to the nerve cells is required. Therefore, human neural stem cells efficient neural differentiation induction, in order to implement a reliable and simple method of development has been desired and, also, be molecular understanding of the mechanism has been desired.
For example, human neural stem cells to differentiate into neuronal cells from the as a method, in the non-patent document 3, for human neural stem cells from the culture solution, is applied in a stepwise manner between 18, B27 (1x) (20ng/ml) (registered trademark) and BDNF (registered trademark) was added to Neurobasal medium is replaced with a, human neural stem cell derived neuronal differentiation from the reported to be executed. In accordance with the method of induction, about 2 weeks, immature neurons (DCX) marker doublecortin, βIII-Tubulin be observed. However, this neuron and the induction efficiency is about 40%, sufficient induction efficiency can be obtained is far.
Scope of claims (In Japanese)請求の範囲 [請求項1]
 ヒト神経幹細胞から分化誘導処理により神経細胞を作製する方法であって、
 前記ヒト神経幹細胞を、FADを含む神経細胞分化誘導用培地を用いて分化誘導処理する工程を含む、方法。

[請求項2]
 請求項1に記載の方法であって、
 神経突起の伸長が促進された神経細胞を作製するための、方法。

[請求項3]
 請求項1または2に記載の方法であって、
 前記分化誘導処理する工程において、前記FADが前記ヒト神経幹細胞内へ取り込まれるための細胞膜透過処理を含む、方法。

[請求項4]
 請求項3に記載の方法であって、
 前記細胞膜透過処理が、前記神経細胞分化誘導用培地への膜透過促進剤の添加である、方法。

[請求項5]
 請求項4に記載の方法であって、
 前記膜透過促進剤がジメチルスルフォオキシドまたはメタノールである、方法。

[請求項6]
 請求項1~5のいずれか一項に記載の方法であって、
 前記FADが1~100nMの範囲で前記神経細胞分化誘導用培地中に含まれる、方法。

[請求項7]
 請求項4または5に記載の方法であって、
 前記膜透過促進剤が0.01~1%(V/V)の範囲で前記神経細胞分化誘導用培地中に含まれる、方法。

[請求項8]
 請求項1~7のいずれか一項に記載の方法であって、
 前記ヒト神経幹細胞が、体細胞由来、iPS細胞由来、ES細胞由来、胎児脳由来、または、生体脳由来の神経幹細胞である、方法。

[請求項9]
 請求項1~8のいずれか一項に記載の方法であって、
 前記分化誘導処理工程の前に、EGFおよび/またはbFGFの存在下、神経幹細胞を前培養する工程をさらに含む、方法。

[請求項10]
 FADを含む神経細胞分化誘導用試薬。

[請求項11]
 分化した神経細胞の神経突起の伸長を促進させるための、請求項10に記載の神経細胞分化誘導用試薬。

[請求項12]
 FAD、および、膜透過促進剤を含む神経細胞分化誘導用キット。

[請求項13]
 分化した神経細胞の神経突起の伸長を促進させるための、請求項12に記載の神経細胞分化誘導用キット。

  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • NATIONAL INSTITUTE OF ADVANCED INDUSTRIAL SCIENCE AND TECHNOLOGY
  • Inventor
  • HIRANO Kazumi
  • NAMIHIRA Masakazu
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH BN BR BW BY BZ CA CH CL CN CO CR CU CZ DE DJ DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JO JP KE KG KH KN KP KR KW KZ LA LC LK LR LS LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PA PE PG PH PL PT QA RO RS RU RW SA SC SD SE SG SK SL SM ST SV SY TH TJ TM TN TR TT TZ UA UG US UZ VC VN ZA ZM ZW
ARIPO: BW GH GM KE LR LS MW MZ NA RW SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ RU TJ TM
EPO: AL AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LT LU LV MC MK MT NL NO PL PT RO RS SE SI SK SM TR
OAPI: BF BJ CF CG CI CM GA GN GQ GW KM ML MR NE SN ST TD TG
Reference ( R and D project ) Biomedical Research Institue,Molecular Neurophysiology Group

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