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High-throughput preparation method of three-dimensional hydrogel array for drug screening

A hydrogel, high-throughput technology, applied in biochemical equipment and methods, specific-purpose bioreactors/fermenters, microorganisms, etc., can solve problems such as drug failure, achieve label-free, convenient operation, and easy operation quick effect

Pending Publication Date: 2021-11-16
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Traditional 2D drug screening platforms often fail in animal or clinical trials

Method used

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  • High-throughput preparation method of three-dimensional hydrogel array for drug screening
  • High-throughput preparation method of three-dimensional hydrogel array for drug screening
  • High-throughput preparation method of three-dimensional hydrogel array for drug screening

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preparation example Construction

[0041] Such as figure 1 As shown, the high-throughput preparation method of the three-dimensional hydrogel array for drug screening provided by the embodiment of the present disclosure includes the following steps:

[0042] Step S1: preparing a superhydrophobic non-adherent cell culture interface;

[0043] Step S2: Prepare a three-dimensional extracellular matrix-mimicking hydrogel that simulates the cell microenvironment, and use the superhydrophobic non-adherent cell culture interface to suspend the three-dimensional extracellular matrix-mimicking three-dimensional hydrogel with cells to be tested in high throughput In culture medium, a suspended three-dimensional hydrogel array is formed.

[0044] According to an embodiment of the present disclosure, a superhydrophobic non-adherent cell culture interface is prepared as described in step S1, and the superhydrophobic non-adherent interface has the following two elements: (a) superhydrophobic; (b) low adhesion. Among them, s...

example 1

[0066] Example 1: PDMS coating method. The present disclosure preferably recommends using the PDMS method. Experiments have proved that the ratio of PDMS precursor to cross-linking agent will not affect the final drug screening results. The present disclosure recommends using a mass ratio of PDMS precursor to cross-linking agent between 1:10 and 1:30. Use a homogenizer to mix the PDMS precursor and cross-linking agent evenly. After defoaming, use a homogenizer to evenly coat it on the surface of the target interface. Attached cell culture interface.

example 2

[0067] Example 2: Pluronic self-assembly method. Prepare a PBS solution containing 0.2% Pluronic by mass, and soak it on the surface of the hydrophilic material. Pluronic is a hydrophilic-hydrophobic-hydrophilic triblock polymer, and its hydrophilic segment self-assembles with the hydrophilic surface, exposing the hydrophobic propylene oxide segment on the substrate surface and inhibiting cell adhesion. Taking F127 as an example, a stock solution of F127 with a mass percentage of 2% can be prepared in advance and diluted to 0.2% when used. Add the F127 working solution to the well plate or petri dish, let it stand for at least 1 hour, wash it thoroughly with DI water, dry it, and then sterilize it with ultraviolet radiation to obtain a super-hydrophobic non-adherent cell culture interface.

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Abstract

The invention discloses a high-throughput preparation method of a three-dimensional hydrogel array for drug screening. The high-throughput preparation method comprises the following steps: preparing a super-hydrophobic non-adhesive cell culture interface; preparing the simulated extracellular matrix three-dimensional hydrogel for simulating a cell microenvironment: suspending the simulated extracellular matrix three-dimensional hydrogel for three-dimensionally culturing cells to be detected in a culture medium in a high-flux manner by utilizing the super-hydrophobic non-adhesive cell culture interface to form a suspended three-dimensional hydrogel array. According to the present invention, by using the super-hydrophobic non-adhesion interface, the three-dimensional hydrogel for three-dimensional culture of the to-be-detected cells is suspended in the culture medium in a high flux manner to form the array. The area shrinkage rate of the three-dimensional hydrogel is adopted as the read-out result of drug screening, such that the high sensitivity is provided for the drug screening using the cell shrinkage force as the target point. The method has the advantages of high flux, low cell consumption, no need of labeling and real-time dynamic analysis.

Description

technical field [0001] The present disclosure relates to the technical field of high-throughput drug screening, in particular to a high-throughput preparation method of a three-dimensional hydrogel array for drug screening. Background technique [0002] The application of three-dimensional cultured microtissues in vitro to the drug screening system is closer to the real situation in vivo than the traditional drug screening platform that uses biochemical, genetic, and single-cell analysis as drug screening indicators. Traditional 2D drug screening platforms often fail in animal or clinical trials. Although the drug screening platform based on tissue engineering cannot completely replace drug tracking in animals, it is very important for reducing the cost of experiments, shortening the research cycle and reducing the consumption of experimental animals. High-content drug screening (HCS) can display the average drug response of a large number of cells, which has unique advanta...

Claims

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Application Information

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IPC IPC(8): C12N5/02C12M3/02C12M1/22
CPCC12N5/0062C12M23/10C12M25/14C12N2513/00C12N2533/90C12N2533/54C12N2533/52C12N2533/56Y02A50/30
Inventor 熊春阳张青
Owner PEKING UNIV
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