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

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...

AI Technical Summary

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

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