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

Foreign code F110005812
File No. K02807WO
Posted date Oct 19, 2011
Country United States of America
Application number 200913128360
Gazette No. 20110217726
Gazette No. 8828679
Date of filing Nov 10, 2009
Gazette Date Sep 8, 2011
Gazette Date Sep 9, 2014
International application number JP2009069124
International publication number WO2010055829
Date of international filing Nov 10, 2009
Date of international publication May 20, 2010
Priority data
  • P2008-289011 (Nov 11, 2008) JP
  • 2009WO-JP69124 (Nov 10, 2009) WO
Title Method and kit for detecting biological signal of three-dimensional cell culture material
Abstract (US8828679)
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.
Scope of claims [claim1]
1. A method for detecting a biological signal of a construct that is produced by a layer-by-layer (LBL) method, comprising: providing a construct that is produced by an LBL method and contains at least two cell layers, an extracellular matrix, and a sensor particle capable of detecting a biological signal; and
observing the sensor particle optically,
wherein the construct is provided so that the extracellular matrix is formed between the cell layers, thereby laminating the cell layers to one another, and
wherein the construct is provided so that 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 claim 1, 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 1, wherein the 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 a first solution and a second solution 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 cell layers,
wherein a combination of an ingredient of the first solution and an ingredient of the second solution is 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.
[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 method for evaluating a construct that is produced by a layer-by-layer (LBL) method, comprising: detecting a biological signal using the method according claim 1; and
analyzing a cellular activity based on a detection result of the biological signal.
[claim8]
8. A method for evaluating a test substance with respect to an organism, comprising: bringing a construct that is produced by a layer-by-layer (LBL) method 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 construct using the method according to claim 1.
  • Inventor, and Inventor/Applicant
  • MATSUSAKI MICHIYA
  • AKASHI MITSURU
  • OKANO KAZUNORI
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
IPC(International Patent Classification)
Reference ( R and D project ) PRESTO Structures and control of interfaces AREA
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