Device for preparing cell cluster, and construction method and application of device

A cell and cluster technology, used in tissue cell/virus culture devices, biochemical equipment and methods, biochemical instruments, etc., can solve the problem that the size and shape of the micropores cannot meet the custom requirements, and it is difficult to obtain a large number of cell clusters. , quantity, size and other problems, to achieve the effect of controllable physical and chemical properties, avoiding the reversal of yin and yang molds, and controllable performance

Active Publication Date: 2022-01-14
TSINGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

By constructing the substrate on a conventional porous culture dish, a soft bottom that is conducive to cell clustering and culture can be provided, but the number and size are limited, and it is difficult to obtain a large number of uniform cell clusters; commercial microporous products can meet the quantity and uniform size requirements, but the general product is still a plastic substrate, the rigidity is too high, and the size and shape of the micropores cannot meet the custom requirements, and cannot be reused, and the cost and expense are too high

Method used

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  • Device for preparing cell cluster, and construction method and application of device
  • Device for preparing cell cluster, and construction method and application of device
  • Device for preparing cell cluster, and construction method and application of device

Examples

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preparation example Construction

[0078] The present invention provides a preparation method of the above cell cluster preparation device. The device preparation method comprises the following steps:

[0079] 1) Using etching or photolithography to fabricate a template with micro-pattern cavities arranged in an array;

[0080] 2) Using the mold forming method, inject liquid or semi-solid overmolding material into the template, and peel it off from the template after solidification and molding to obtain the overmolding stamp;

[0081] 3) Mix the base material with the cross-linking agent (perform a vacuum operation to remove air bubbles), place the mold stamp in the above mixture, and solidify and form;

[0082] 4) peeling off the overmolding stamp from the above-mentioned molding material, or removing the overmolding stamp from the above-mentioned molding material by chemical or physical methods, to obtain a substrate with micro-pattern cavities arranged in an array;

[0083]5) Fix the substrate in the micro...

Embodiment 1

[0121] Example 1 The method of making a cell cluster preparation device with a degradable overturned mold stamp

[0122] 1. Preparation of micrographic templates

[0123] The micrographic template is made of photolithographic polymethyl methacrylate to prepare a circular template with an overall diameter of 3 cm. The template is an inverted cone-shaped array containing 1500 micro cavities. The distance between adjacent cavities is 200 μm. The figure is an inverted cone with a square upper surface, and the upper surface area is 0.25mm 2 , with a depth of 500 μm.

[0124] 2. Preparation of biodegradable stamps

[0125] The mold stamp is made of agar material. Agar solution with a concentration of 5% is used, dissolved under high temperature and high pressure, poured into the micrographic template while it is hot, and 2ml of agar is poured each time, and vacuum is used to remove air bubbles before the agar solution is solidified. After the agar has cooled and solidified, care...

Embodiment 2

[0132] Example 2 The method of making a cell cluster preparation device with a non-degradable mold stamp

[0133] 1. Preparation of micrographic templates

[0134] The micrographic template is made of etched polymethyl methacrylate to prepare a circular template with an overall diameter of 3 cm. The template is an inverted cone-shaped array containing 1500 micro cavities, and the distance between adjacent cavities is 200 μm. The figure is an inverted cone with a square upper surface, and the upper surface area is 0.25mm 2 , with a depth of 500 μm.

[0135] 2. Preparation of non-degradable flip stamps

[0136] The non-degradable flip stamp is made of silicone material. Mix the silica gel matrix (the main components are vinyl silicone oil and silicon dioxide) with the cross-linking agent (the main component is hydrogen-containing silicone oil) at a volume ratio of 1:1, and quickly pour it onto the template and press it hard to ensure that the material completely covers the mi...

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Abstract

The invention provides a device for preparing a cell cluster, and a construction method and application of the device. The device comprises a cell culture plate and a substrate which is fixed in the micropores of the cell culture plate and is provided with micro-pattern cavities arranged in an array form. Each micro-pattern cavity forms a space for growth of the cell cluster, and the shape and the volume of the micro-pattern cavity can regulate and control the property of the obtained cell cluster. The preparation method comprises the following steps of preparing a size-adjustable micro-pattern template, and carrying out two times of mold turning to obtain the array-arranged micro-pattern substrate. The device provided by the invention can be used for conveniently preparing cell clusters on a large scale and can be repeatedly used; and the elastic modulus of the substrate can be adjusted so as to meet the requirements of co-culture of various different cells. The cell cluster prepared by the device is high in activity, has a uniform size and complete shape, is controllable in performance and good in biological performance, and can be used for cell therapy, high-throughput model construction, multi-cell co-culture, stem cell differentiation, organoid construction, drug screening, tissue regeneration, in-vitro artificial systems and the like.

Description

technical field [0001] The invention relates to the technical fields of biotechnology and tissue engineering, in particular to a device for preparing cell clusters and its construction method and application. Background technique [0002] The traditional cell culture method is planar culture. The monolayer cell culture obtained by planar culture is different from the physiological structure of cells in vivo, which will affect the survival and function of cells. Using self-assembly between cells to form cell clusters, a quasi-3D environment can be constructed. Compared with planar culture, three-dimensional cell culture can reconstruct the interaction between cells and cells, and between cells and matrix, and reduce the difference between in vitro culture and native tissue. For example, cells such as embryonic stem cells, mesenchymal stem cells, and various tumor cells have the characteristic of clustering. In addition, islet β cells need to form cell clusters to maintain t...

Claims

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Application Information

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IPC IPC(8): C12M3/00C08L83/04C08L89/00C08J5/18C08J3/24
CPCC12M23/12C12M23/20C08J5/18C08J3/24C08J2383/04C08J2489/00C08J2483/04C08J2389/00C08L83/04C12M3/00C08L89/00
Inventor 姚睿苏奕君
Owner TSINGHUA UNIV
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