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Cell culture substratum, method for producing cell-containing material, method for producing cell culture substratum, method for observing cells, and cell culture substratum maintenance liquid

By forming multiple concave and convex structures on the cell culture substrate and coating them with inorganic materials, combined with proteins, the cell adhesion rate and survival rate are improved, and the low adhesion rate, survival rate, and insufficient thermal stability of polystyrene cell culture dishes are solved. issues, achieving durability and recycling.

Active Publication Date: 2019-01-04
OHARA +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, when polystyrene cell culture dishes are used as culture substrates, there are problems of low adhesion rate and cell viability of cultured cells
In addition, since polystyrene cell culture dishes have low heat resistance, there is a problem that they cannot be recycled by heat sterilization

Method used

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  • Cell culture substratum, method for producing cell-containing material, method for producing cell culture substratum, method for observing cells, and cell culture substratum maintenance liquid
  • Cell culture substratum, method for producing cell-containing material, method for producing cell culture substratum, method for observing cells, and cell culture substratum maintenance liquid
  • Cell culture substratum, method for producing cell-containing material, method for producing cell culture substratum, method for observing cells, and cell culture substratum maintenance liquid

Examples

Experimental program
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Effect test

preparation example Construction

[0107] (Preparation method of cell contents)

[0108] The method for producing a cell content according to the present embodiment includes a step of applying cells to a concavo-convex structure in a cell culture substrate and culturing them. By applying cells to the concavo-convex structure, cell culture can be highly activated, and cell contents such as cell tissue or living tissue having good adhesion to the matrix can be produced.

[0109] In the preparation method of the cell content, cells are cultured using a cell culture substrate made of polystyrene, and the culture time at which the density of the cells is 90% of the culture surface of the cell culture substrate made of polystyrene is defined as 1T. In some cases, it is preferable to culture cells for a culture time of 1 / 2T to 2 / 3T using the cell culture substrate of this embodiment. For example, when the culture time for 90% of the culture surface of a cell culture substrate made of polystyrene is 24 hours, by using...

Embodiment 1

[0121] Both surfaces of the quartz glass substrate made of silica were polished using a double-side grinder (manufactured by Hamai Sangyo Co., Ltd., 4-way grinding disc). In the grinding process, zirconia-based alumina (AZ) powder manufactured by Heisei Sankei Co., Ltd. 2 , Platen rotation speed 20 ~ 50rpm implementation. Both surfaces of the polished quartz glass substrate were polished using a double-side polisher (manufactured by Hamai Sangyo Co., Ltd., 4-way polishing disc). In the polishing process, a polyurethane foam pad is used as a polishing pad, and MIREK E manufactured by Mitsui Metal Mining Co., Ltd. 2 , Platen rotation speed 20 ~ 50rpm implementation. Accordingly, a cell culture substrate composed of a silica substrate was prepared.

Embodiment 2

[0123] (Preparation method of a cell culture substrate in which a silica substrate is coated with a porous silica film)

[0124] Add 524 mg of cetyltrimethylammonium chloride (CTAC) to 1.4 ml of water, stir until dispersed, and obtain solution A. Then, 0.472 ml of water and 0.1 ml of 0.1N-HCl were added to 1.94 ml of orthomethyl silicate (TMOS), and stirred at 30° C. for 60 minutes to obtain a solution B. Solution A and solution B were mixed so that the molar ratio of TMOS / CTAC was 8, and 0.1 ml of 1N-HCl was further added to obtain mixed solution C. After the mixture C was stirred at room temperature (20° C.) for 15 minutes, spin coating was performed at 6000 rpm for 10 seconds on the silica substrate (12.5 mm × 25 mm) of Example 1 having a concavo-convex shape on the culture surface. . The silicon dioxide substrate coated with the mixed solution C was heated at 60° C. for 18 hours to form a CTAC / TMOS composite film. In addition, the silica substrate on which the composite...

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Abstract

The purpose of the invention is to provide a cell culture substratum which has excellent resistance to liquid culture media and low cytotoxicity, can achieve a high cell adhesion ratio and a high viability of cultured cells, has excellent thermal stability, and is less likely to absorb ultraviolet ray. A cell culture substratum which is provided with a substrate made from an inorganic material andhas multiple concavo-convex structures on a culturing surface thereof, wherein, when the concavo-convex structures are measured with an atomic force microscope in accordance with JISB0601 and JISR1683 (measured area: a 1 [mu]m-square, cut-off value of a low-pass contour curve filter: 1 nm, cut-off value of a high-pass contour curve filter: 170 nm), the average of the lengths of contour curve elements of the concavo-convex structures is 1 to 170 nm as measured in at least one direction (when a curve showing long-wavelength components that are blocked by the high-pass contour curve filter is converted to a straight line by the least square method, the average line is a line that is parallel with the straight line and indicates a height cumulative relative frequency distribution in the contour curve of 50%).

Description

technical field [0001] The invention relates to a cell culture substrate, a preparation method of cell contents, a preparation method of the cell culture substrate, a cell observation method and a maintenance solution of the cell culture substrate. Background technique [0002] In recent years, regenerative medicine technology that transplants cells and tissues obtained by culturing cells in vitro into the body has attracted attention. For example, the research and development of regenerative medicine technology that regenerates stem cells such as iPS cells and living tissues such as bones, blood vessels, skin, and heart by culturing various cells differentiated into living tissues in cell culture dishes has made great progress. [0003] As a substrate for cell culture, polystyrene cell culture dishes are generally used. However, when a polystyrene cell culture dish is used as a culture substrate, there is a problem that the adhesion rate and cell survival rate of cultured ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00C12M1/34C12N1/00C12N5/07
CPCC12M1/34C12M3/00C12N1/00C12M23/20C12N5/0654C12N2533/12C12N2533/18C12N2535/00C12N5/0068C12M23/02C12N2533/10C12N2533/00C12N2533/14
Inventor 犬井正彦茶谷直多贺谷基博本塚智
Owner OHARA
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