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MICROFLUID DEVICE AND THREE-DIMENSIONAL MICROCULTURE METHOD FOR CELL

Foreign code F180009647
File No. 4484
Posted date Nov 21, 2018
Country WIPO
International application number 2015JP055178
International publication number WO 2015129673
Date of international filing Feb 24, 2015
Date of international publication Sep 3, 2015
Priority data
  • P2014-034166 (Feb 25, 2014) JP
Title MICROFLUID DEVICE AND THREE-DIMENSIONAL MICROCULTURE METHOD FOR CELL
Abstract The present invention provides a microfluid device comprising a cell culture chamber connected to at least two openings. When cells and hydrogel are introduced to the cell culture chamber and cells are cultured in a three-dimensional gel medium, biologically active substances can be fed from at least one opening to the cell culture chamber while a concentration gradient is formed inside the chamber.
Outline of related art and contending technology BACKGROUND ART
Cells, the extracellular in vivo microenvironment under its functions in control is being performed. The extracellular microenvironment, mainly (i) growth factor, vitamin, gas molecules such as soluble factors, extracellular matrix proteins (ii), such as the hardness of the non-soluble factors, cell-cell interactions (iii), is configured from. These factors is complicated, and while the tightly controlled, cell function control is performed. That is, human ES cells and human iPS cells for the purpose of cell function in order to freely control, this extracellular microenvironment as it is necessary to freely control.
However, these control operations are performed in a micrometer scale and very small, conventional culturing of the cells, experimental method, using a culture dish or plate 2 is three-dimensional environment, cannot be reproduced. Therefore, was difficult by a conventional method, 3-dimensional cell culture, can be made to the experimental environment has been demanded.
Culturing the cell in the conventional human ES/iPS, experimental method, such as three-dimensional culture dish was performed in a simulated environment 2 (non-patent document 1, 2).
However, the original, 3 cells are placed in a three-dimensional environment, 2 expression and the function of the three-dimensional environment cannot be. Human ES/iPS cells were used in tissue engineering, the three-dimensional environment for 3 is very important.
The size and the very important factors. Cells in vivo, in micrometer scale controlled micro-environment. For example, the concentration gradient and of a soluble factor, the hardness of the extracellular matrix and the like. These factors by a conventional method was difficult to control. Of course, these factors can be comprehensively analyzed, was almost impossible.
Scope of claims (In Japanese)請求の範囲 [請求項1]
少なくとも2つの開口部と接続した細胞培養チャンバーを有し、前記細胞培養チャンバー内に細胞とヒドロゲルを導入して三次元ゲル培地で培養したときに、少なくとも1つの開口部から細胞培養チャンバーへの生理活性物質の供給を前記チャンバー内での濃度勾配を形成しながら行うことができる、マイクロ流体デバイス。

[請求項2]
前記ヒドロゲルは細胞培養チャンバー内で形成される、請求項1に記載のマイクロ流体デバイス。

[請求項3]
少なくとも1つの開口部と細胞培養チャンバーとを接続する流路が前記チャンバーの径と比較して細いことを特徴とする、請求項1又は2に記載のマイクロ流体デバイス。

[請求項4]
請求項1~3のいずれかに記載のマイクロ流体デバイスの細胞培養チャンバーに細胞と流動状ヒドロゲルを導入する工程、前記ヒドロゲルをゲル状に変換する工程、少なくとも1つの開口部から生理活性物質を細胞培養チャンバー内で濃度勾配を形成するように供給して前記生理活性物質の存在下に細胞を培養する工程を含む、細胞の微小3次元培養法。

[請求項5]
前記細胞が多能性幹細胞である、請求項4に記載の細胞の微小3次元培養法。

[請求項6]
前記細胞がヒト多能性幹細胞である、請求項4に記載の細胞の微小3次元培養法。

[請求項7]
複数の生理活性物質を濃度勾配を形成するように細胞培養チャンバーに供給することを特徴とする、請求項4~6のいずれかに記載の微小3次元培養法。

  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • KYOTO UNIVERSITY
  • Inventor
  • KAMEI, Kenichiro
  • CHEN, Yong
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 DK DM DO DZ EC EE EG ES FI GB GD GE GH GM GT HN HR HU ID IL IN IR IS JP KE KG KN KP KR 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
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