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Method and kit for detecting biological signal of three-dimensional cell culture material

外国特許コード F110005792
整理番号 K02807WO
掲載日 2011年9月20日
出願国 欧州特許庁(EPO)
出願番号 09826077
公報番号 2357251
公報番号 2357251
出願日 平成21年11月10日(2009.11.10)
公報発行日 平成23年8月17日(2011.8.17)
公報発行日 平成26年9月24日(2014.9.24)
国際出願番号 JP2009069124
国際公開番号 WO2010055829
国際出願日 平成21年11月10日(2009.11.10)
国際公開日 平成22年5月20日(2010.5.20)
優先権データ
  • 2009JP069124 (2009.11.10) WO
  • 特願2008-289011 (2008.11.11) JP
発明の名称 (英語) Method and kit for detecting biological signal of three-dimensional cell culture material
発明の概要(英語) Disclosed is a method for detecting a biological signal of a three-dimensional cell culture construct.
The method for detecting a biological signal of a three-dimensional cell culture construct includes: providing a three-dimensional cell culture construct that contains at least two cell layers laminated to each other and a sensor particle capable of detecting a biological signal; and observing the sensor particle optically.
Preferably, the three-dimensional cell culture construct contains an extracellular matrix including a combination of a protein or polymer having an RGD sequence and a protein or polymer that interacts with the protein or polymer having the RGD sequence, or a combination of a protein or polymer that is positively charged and a protein or polymer that is negatively charged.(see diagramm)
特許請求の範囲(英語) [claim1]
1. A method for detecting a biological signal of a three-dimensional cell culture construct that contains at least two cell layers laminated to each other, an extracellular matrix, and a sensor particle capable of detecting a biological signal, comprising: optically observing the sensor particle in said three-dimensional cell culture construct, wherein the extracellular matrix is formed at least between the cell layers and the sensor particle is placed in the extracellular matrix between the cell layers.
[claim2]
2. The method according to claim 1, wherein the sensor particle contains a substance having a sensing function, a carrier for supporting the substance having a sensing function, and a biocompatible basic polymer layer and a biocompatible acidic polymer layer that are laminated alternately on a surface of the carrier, and the carrier is a porous particle.
[claim3]
3. The method according to claim 1, wherein the optical observation of the sensor particle includes visualization and/or numeric conversion of the biological signal.
[claim4]
4. The method according to any one of claims 1 to 3, wherein the extracellular matrix includes a combination of a protein or polymer having an RGD sequence and a protein or polymer that interacts with the protein or polymer having the RGD sequence, or a combination of a protein or polymer that is positively charged and a protein or polymer that is negatively charged.
[claim5]
5. The method according to claim 4, wherein the three-dimensional cell culture construct is capable of being produced by a process, comprising: forming a cell layer by introducing a cell-containing solution; forming an extracellular matrix by introducing (i) a solution comprising a protein or polymer having an RGD sequence and a solution comprising a protein or polymer that interacts with the protein or polymer having the RGD sequence alternately, or (ii) a solution comprising a protein or polymer that is positively charged and a solution comprising a protein or polymer that is negatively charged alternately; forming the extracellular matrix and the cell layer alternately to laminate the cell layers; and placing the sensor particle capable of detecting a biological signal between the cell layers, wherein the combination of the proteins or polymers of the extracellular matrix forming solutions provides said combination of a protein or polymer having an RGD sequence and a protein or polymer that interacts with the protein or polymer having the RGD sequence, or said combination of a protein or polymer that is positively charged and a protein or polymer that is negatively charged.
[claim6]
6. The method according to claim 1, wherein the sensor particle contains a substance having a sensing function, a carrier for supporting the substance having a sensing function, and a basic polymer layer and an acidic polymer layer that are laminated alternately on a surface of the carrier.
[claim7]
7. A detection kit for use in the method according to any one of claims 1 to 6, including a three-dimensional cell culture construct that contains at least two cell layers laminated to each other, an extracellular matrix formed at least between the cell layers, and a sensor particle capable of detecting a biological signal placed in the extracellular matrix formed between the cell layers.
[claim8]
8. The detection kit according to claim 7, wherein the sensor particle contains a substance having a sensing function, a carrier for supporting the substance having a sensing function, and a biocompatible basic polymer layer and a biocompatible acidic polymer layer that are laminated alternately on a surface of the carrier.
[claim9]
9. A method for producing a three-dimensional cell culture construct, comprising: forming a cell layer by introducing a cell-containing solution; forming an extracellular matrix by introducing (i) a solution comprising a protein or polymer having an RGD sequence and a solution comprising a protein or polymer that interacts with the protein or polymer having the RGD sequence alternately, or (ii) a solution comprising a protein or polymer that is positively charged and a solution comprising a protein or polymer that is negatively charged alternately; forming the extracellular matrix and the cell layer alternately to laminate the cell layers; and placing a sensor particle capable of detecting a biological signal between the cell layers, wherein the combination of proteins or polymers of the extracellular matrix forming solutions provides said combination of a protein or polymer having an RGD sequence and a protein or polymer that interacts with the protein or polymer having the RGD sequence, or said combination of a protein or polymer that is positively charged and a protein or polymer that is negatively charged.
[claim10]
10. A method for evaluating a three-dimensional cell culture construct, comprising: detecting a biological signal using the method according to any one of claims 1 to 6; and analyzing a cellular activity based on a detection result of the biological signal.
[claim11]
11. An in vitro method for evaluating how a test substance influences an organism, comprising: bringing a three-dimensional cell culture construct which is a tissue model for said organism into contact with a test substance selected from the group consisting of compounds, medical compositions, cosmetics and foods; and detecting a biological signal of the three-dimensional cell culture construct using the method according to any one of claims 1 to 6.
  • 出願人(英語)
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
  • 発明者(英語)
  • MATSUSAKI MICHIYA
  • AKASHI MITSURU
  • OKANO KAZUNORI
国際特許分類(IPC)
欧州特許分類/主・副
  • G01N033/50D2E8
  • G01N033/50D2J4
  • G01N033/543D
指定国 Contracting States: AT BE BG CH CY CZ DE DK EE ES FI FR GB GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO PL PT RO SE SI SK SM TR
参考情報 (研究プロジェクト等) PRESTO Structures and control of interfaces AREA
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