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CULTURE METHOD AND CULTURE DEVICE

Foreign code F120006970
File No. S2010-1166-C0
Posted date Oct 29, 2012
Country WIPO
International application number 2011JP070533
International publication number WO 2012033181
Date of international filing Sep 8, 2011
Date of international publication Mar 15, 2012
Priority data
  • P2010-203741 (Sep 10, 2010) JP
Title CULTURE METHOD AND CULTURE DEVICE
Abstract Provided are a culture method and a culture device for efficiently culturing cells and/or a cell tissue suitable for medical uses. The culture method, whereby cells adhered to an electrode layer (30) are cultured, comprises: a first step for adhering the cells to the electrode layer (30) and culturing the same, wherein said electrode layer (30)is coated with an oligopeptide (10) having a terminal amino acid (11) constituting one end thereof and being capable of binding to the electrode layer via thiolate, a cell-adhesive sequence (14) constituting the other end thereof and comprising a cell-adhesive amino acid sequence, and an alternate sequence (13) binding to the aforesaid one end of the cell-adhesive sequence (14) and consisting of a plurality of acidic amino acids (13a) and a plurality of basic amino acids (13b), said acidic amino acids and basic amino acids being alternately bonded, one by one, to each other; and a second step for applying, to the electrode layer (30) carrying the cells adhered thereto, a potential capable of inducing the reductive elimination of the oligopeptide (10) and thus eliminating the cells from the electrode layer (30).
Outline of related art and contending technology BACKGROUND ART
Conventional, is adhered to the surface of the substrate as a method of recovering the cultured cells, for example, a method using a proteolytic enzyme and a chelating agent, a method using a magnetic force, a method using a temperature-responsive polymer, is a method of using a photoresponsive polymer.
However, the use of proteolytic enzymes and a chelating agent in the method, cell injury was received. In addition, in the method using a magnetic force necessary to magnetize cells are. The temperature-responsive polymer or a photoresponsive polymer used in this method through the culture period was necessary to control the temperature and light.
Therefore, in Patent Document 1, via thiolate spacer substance is bound to the electrode surface to adhere the cultured cells, then, the electrode has been adhered to the cell surface, the spacer material by applying a potential to reduction desorption, separated from the surface of the electrode into the cells has been proposed a method.
In addition, in non-patent document 1, the center of the cell adhesive domain (RGD), which has cysteine residues at both ends of the oligopeptide via a gold adhered cells onto the surface, then, by reduction desorption into the cells separated from the surface of the gold has been proposed a method.
Scope of claims (In Japanese)請求の範囲 [請求項1]
 電極層に接着した細胞を培養する方法であって、
 一方の末端を構成しチオレートを介して前記電極層に結合する末端アミノ酸と、
 他方の末端を構成し細胞接着性のアミノ酸配列からなる細胞接着性配列と、
 前記細胞接着性配列の前記一方の末端側に結合し複数の酸性アミノ酸と複数の塩基性アミノ酸とが1つずつ交互に結合してなる交互配列と、
 を有するオリゴペプチドで被覆された前記電極層に前記細胞を接着させて培養する第一工程と、
 前記細胞が接着している前記電極層に、前記オリゴペプチドが還元脱離する電位を印加して、前記細胞を前記電極層から脱離させる第二工程と、
 を含む
 ことを特徴とする培養方法。

[請求項2]
 前記電極層は、培養液中に浮いた状態で保持される多孔質膜の表面に形成されている
 ことを特徴とする請求項1に記載の培養方法。

[請求項3]
 前記第一工程において、前記電極層に前記細胞を接着させて培養することで前記電極層に接着した細胞組織体シートを形成し、
 前記第二工程において、前記細胞組織体シートが接着している前記電極層に、前記オリゴペプチドが還元脱離する前記電位を印加して、前記細胞組織体シートを前記電極層から脱離させる
 ことを特徴とする請求項1又は2に記載の培養方法。

[請求項4]
 前記電極層から脱離させた複数の前記細胞組織体シートを積層して、前記細胞組織体シートの積層体を形成する第三工程をさらに含む
 ことを特徴とする請求項3に記載の培養方法。

[請求項5]
 細胞を接着させて培養するための電極層を有する培養装置であって、
 前記電極層は、
 一方の末端を構成しチオレートを介して前記電極層に結合する末端アミノ酸と、
 他方の末端を構成し細胞接着性のアミノ酸配列からなる細胞接着性配列と、
 前記細胞接着性配列の前記一方の末端側に結合し複数の酸性アミノ酸と複数の塩基性アミノ酸とが1つずつ交互に結合してなる交互配列と、
 を有するオリゴペプチドで被覆されている
 ことを特徴とする培養装置。

[請求項6]
 前記電極層は、培養液中に浮いた状態で保持される多孔質膜の表面に形成されている
 ことを特徴とする請求項5に記載の培養装置。

  • Applicant
  • ※All designated countries except for US in the data before July 2012
  • UNIVERSITY OF TSUKUBA
  • Inventor
  • FUKUDA, Junji
  • SUZUKI, Hiroaki
  • MOCHIZUKI, Naoto
  • KAKEGAWA, Takahiro
IPC(International Patent Classification)
Specified countries National States: AE AG AL AM AO AT AU AZ BA BB BG BH 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 IS JP KE KG KM KN KP KR KZ LA LC LK LR LS LT LU LY MA MD ME MG MK MN MW MX MY MZ NA NG NI NO NZ OM PE PG PH PL PT QA RO RS RU RW 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 SD SL SZ TZ UG ZM ZW
EAPO: AM AZ BY KG KZ MD 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 ML MR NE SN TD TG
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