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Hemispherical micropore array substrate for three-dimensional cell sphere culture and preparation method of hemispherical micropore array substrate

A three-dimensional cell and microwell array technology, applied in general culture methods, cell culture supports/coatings, biochemical equipment and methods, etc. Collect difficulties and other problems, achieve the effect of simple operation, enhanced collision between cells, and low cost

Pending Publication Date: 2021-08-06
SOUTH CHINA UNIV OF TECH
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  • Description
  • Claims
  • Application Information

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Problems solved by technology

In the prior art, microsphere arrays are also used to prepare PDMS arc-shaped culture substrates, but what they obtain is spherical cavities with small openings, and the signal transmission between cells cannot be carried out well, and it is difficult to collect cell spheres; at the same time, because PDMS is relatively Hydrophobic must undergo a series of pretreatments before use, and the low cell capture rate often requires high cell density to form aggregates

Method used

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  • Hemispherical micropore array substrate for three-dimensional cell sphere culture and preparation method of hemispherical micropore array substrate
  • Hemispherical micropore array substrate for three-dimensional cell sphere culture and preparation method of hemispherical micropore array substrate
  • Hemispherical micropore array substrate for three-dimensional cell sphere culture and preparation method of hemispherical micropore array substrate

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preparation example Construction

[0044] A preparation method for a hemispherical microwell array substrate for three-dimensional cell spheroid culture, comprising the following steps (refer to figure 1 shown):

[0045] (1) Stick a layer of double-sided adhesive on the surface of the glass, and use the double-sided adhesive to stick the stainless steel screen with a mesh size of 30 mesh on the surface to obtain a sandwich structure of double-sided adhesive-glass-stainless steel screen. Pour excess 400μm stainless steel microspheres into the sieve, capture a single steel ball in a single sieve micropore by scraping, fix a steel ball in each sieve hole, press, so that the steel ball is pasted by double-sided adhesive, and then The stainless steel screen was removed to obtain a plane with microsphere arrays (the front side of which is neatly arranged microsphere arrays).

[0046] (2) Prepare a clean glass substrate (thickness 20mm), apply a layer of UV glue on it with a glue leveler at a speed of 2000rpm, then p...

Embodiment 4

[0059] A preparation method for a hemispherical microwell array substrate for three-dimensional cell spheroid culture, comprising the following steps (refer to figure 1 shown):

[0060] (1) Stick a layer of double-sided adhesive on the surface of the glass, and use the double-sided adhesive to stick the stainless steel screen with a mesh size of 50 mesh on the surface to obtain a sandwich structure of double-sided adhesive-glass-stainless steel screen. Pour excess 300μm stainless steel microspheres into the sieve, capture a single steel ball in a single sieve micropore by scraping, fix a steel ball in each sieve hole, press it so that the steel ball is pasted by double-sided adhesive, and then Pull off the stainless steel screen to obtain a plane with microsphere arrays (the front side is a neatly arranged microsphere array).

[0061] (2) Prepare a clean glass substrate (thickness 20mm), apply a layer of UV glue on it with a glue leveler at a speed of 2000rpm, then press the ...

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Abstract

The invention relates to a hemispherical microarray substrate for three-dimensional cell sphere culture and a preparation method of the hemispherical microarray substrate. The surface of a cell culture substrate disclosed by the invention has a regular hemispherical microarray structure. The method comprises the following steps: pasting a surface adhesive on a plane, pasting a screen on the plane, fixing microspheres in micropores of the screen, and uncovering the screen to obtain the plane with a microsphere array; coating a light-transmitting substrate with ultraviolet glue, pressing the plane with the microsphere array on the ultraviolet glue, performing curing, and separating the light-transmitting substrate from the plane with the microsphere array to obtain a semi-microsphere array template; and adding an agarose solution, performing curing, and separating cured agarose from the microsphere array template to obtain the hemispherical micropore array substrate for three-dimensional cell sphere culture. The hemispherical micropore array structure provided by the method is formed by simple hemispherical array rollover, and is easy to operate, low in cost and free of any complex operation. The cell culture substrate disclosed by the invention can be used for conveniently and massively preparing cell balls with uniform sizes.

Description

technical field [0001] The invention belongs to the field of cell culture, and in particular relates to a hemispherical microwell array substrate for three-dimensional cell aggregate culture and a preparation method thereof. Background technique [0002] The growth method of cells spreading in a single layer on the surface of a certain substrate is called two-dimensional culture, which is a convenient cell culture method for observation and long-term culture. However, with the deepening of the research, people found that the interaction between cells and the cellular microenvironment plays a decisive role in the proliferation, differentiation and function of cells. Obviously, since cells mostly exist in the extracellular matrix in the form of multi-layer cell accumulation in physiological environment, two-dimensional culture cannot well simulate their living environment to meet the research needs. Therefore, how to simulate the in vivo microenvironment of cells has attracte...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/00
CPCC12N5/0068C12N2513/00C12N2533/76
Inventor 杜昶付晓玲萧学敏
Owner SOUTH CHINA UNIV OF TECH
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