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Method for determining undifferentiated state of pluripotent stem cell, method for subculturing pluripotent stem cell, and device for use in said methods

A technology for pluripotent stem cells and cell differentiation, which is applied in the field of determining the undifferentiated state of pluripotent stem cells, and can solve problems such as low data freedom and large-scale measurement equipment

Active Publication Date: 2020-04-10
TOKYO ELECTRON LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

As a method for measuring labeled metabolites, it is generally performed by combining component separation methods such as liquid chromatography (LC), gas chromatography (GC), and capillary electrophoresis (CE) with detection methods such as mass spectrometry (MS) , but sometimes there are cases where the measurement equipment is enlarged and the degree of freedom of data acquisition is low

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  • Method for determining undifferentiated state of pluripotent stem cell, method for subculturing pluripotent stem cell, and device for use in said methods
  • Method for determining undifferentiated state of pluripotent stem cell, method for subculturing pluripotent stem cell, and device for use in said methods
  • Method for determining undifferentiated state of pluripotent stem cell, method for subculturing pluripotent stem cell, and device for use in said methods

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Embodiment 1

[0145] Example 1: Study on the relationship between the undifferentiated state of pluripotent stem cells and the components of the culture medium

[0146] Hereinafter, an example of evaluating the differentiation state of cells by the method of the present invention will be described.

[0147] image 3 It is a schematic diagram showing the implementation procedure for recovering the culture medium sample for evaluation model creation of the cell differentiation state in this example.

[0148] In this example, the human iPS cell line PFX#9 was used. In addition, cells to which a differentiation-inducing stimulus was given to a human iPS cell line were used as test cells, and cells to which a human iPS cell line was not given a differentiation-inducing stimulus (that is, cells maintained in an undifferentiated state) were used as control cells. The procedure from cell culture to analysis of culture medium samples (culture supernatant) in this example will be described below. ...

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Abstract

The present disclosure provides a method for determining the undifferentiated state of a pluripotent stem cell, the method comprising irradiating a culture medium being tested in which the pluripotentstem cell is cultured, with light having wavelengths in all or a part of the range of 190-2500 nm, detecting reflected light, transmitted light, or transmitted and reflected light thereof to obtain absorbance spectral data, and analyzing the absorbances in all or in a partial range of the measured wavelengths of the absorbance spectral data, on the basis of an analysis model which is previously created using a plurality of types of control culture media used in culturing the pluripotent stem cell, to determine the undifferentiated state of the pluripotent stem cell, wherein the plurality of types of control culture media include one or more differentiation-inducing media selected from a medium used in maintaining the undifferentiated state of the pluripotent stem cell, a medium for inducing differentiation to an ectodermal cell, a medium for inducing differentiation to a mesodermal cell, and a medium for inducing differentiation to an endodermal cell.

Description

technical field [0001] The present invention relates to a method for determining the undifferentiated state of pluripotent stem cells, a method for subculture of pluripotent stem cells, and an apparatus used in these methods. Background technique [0002] Pluripotent stem cells are widely used in various fields such as research on tissue differentiation, drug testing, and regenerative medicine due to their pluripotency to differentiate into any tissue. Especially since the establishment of iPS cells, the research in this field has progressed remarkably, and various attempts to realize regenerative medicine are being carried out all over the world. [0003] However, pluripotent stem cells are easy to differentiate, and once differentiated, they may lose their pluripotency. Therefore, the culture of pluripotent stem cells needs to be carried out under the condition of maintaining the undifferentiated state of pluripotent stem cells. One of the most important elements in the c...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/04C12M1/34G01N21/27C12N5/071C12N5/10
CPCG01N21/27C12M41/36C12N5/0696C12N2529/10G01N33/5005C12M31/00C12M41/46C12N5/0607
Inventor 依田祐介畑林邦忠加川健一
Owner TOKYO ELECTRON LTD
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