Preparation and application of high-throughput in-situ observable cell 3D culture plate

A cell culture, high-throughput technology, applied in tissue cell/virus culture devices, biochemical equipment and methods, methods of supporting/immobilizing microorganisms, etc., can solve the problems of poor transparency and limitations of liquid marbles, and achieve effective Facilitate the effect of large-scale production of cells

Pending Publication Date: 2022-01-04
NORTHWESTERN POLYTECHNICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] 1. The preparation of traditional liquid marbles uses powder as the particle source. After the liquid marbles are prepared, the excess powder must be removed, and the liquid marbles must be transferred to a platform such as a petri dish for subsequent operations. This is Qualcomm Massive cell culture and comparative analysis pose significant limitations;
[0010] 2. The surface particles of traditional powder liquid marbles will form clusters of tens of microns or more, resulting in poor transparency of liquid marbles, and it is often necessary to extract the internal substances for observation

Method used

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  • Preparation and application of high-throughput in-situ observable cell 3D culture plate
  • Preparation and application of high-throughput in-situ observable cell 3D culture plate
  • Preparation and application of high-throughput in-situ observable cell 3D culture plate

Examples

Experimental program
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Effect test

Embodiment 1

[0070] The preparation of a high-throughput, in-situ observed cell 3D culture plate comprises the following steps:

[0071] Step 1: Preparation of Alkylated SiO 2 Sol:

[0072] Step 1.1: Measure 16mL of tetraethyl orthosilicate and add it to 200mL of absolute ethanol solution, and keep stirring at room temperature for 20min to completely mix the two to obtain solution A;

[0073] Step 1.2: Add 8.4mL of ammonia water to the solution A obtained in step 1.1. The concentration of ammonia water is 25%-28%. Stir while adding, then continue to stir at room temperature for 40min, let it stand for aging for 7 days, and obtain solution B;

[0074] Step 1.3: Add 4.48 mL of hexamethyldisilazane to the solution B obtained in step 1.2, stir while adding, continue stirring for another 30 minutes, and leave it to age at room temperature for 2 days to obtain alkylated SiO 2 Sol.

[0075] The present invention adopts the ratio of tetraethyl orthosilicate, dehydrated alcohol, ammoniacal liquo...

Embodiment 2

[0086] The preparation of a high-throughput, in-situ observed cell 3D culture plate comprises the following steps:

[0087] Step 1: Preparation of Alkylated SiO 2 Sol, preparation method is with embodiment 1:

[0088] Step 2: cleaning the cell culture plate, the cleaning method is the same as in Example 1;

[0089] Step 3: The specific steps include:

[0090] Step 3.1: Alkylated SiO prepared in step 1.32 Add the sol to each well of the cell culture plate cleaned in step 2, shake gently to make the sol completely cover the inner wall of the cell culture plate hole, and then pour out the sol;

[0091] Step 3.2: Repeat the operation of step 3.1 3 times. When pouring off the excess sol and adding new sol each time, the interval is 10s to make the hole in a semi-wet state, and then blow air with the ear washing ball to make it dry quickly.

[0092] Step 3.3: Repeat the operation of step 3.2 3 times to obtain superhydrophobic SiO with weak binding force on the surface 2 Xerogel-...

Embodiment 3

[0097] The preparation of a high-throughput, in-situ observed cell 3D culture plate comprises the following steps:

[0098] Step 1: Preparation of Alkylated SiO 2 Sol, preparation method is with embodiment 1:

[0099] Step 2: cleaning the cell culture plate, the cleaning method is the same as in Example 1;

[0100] Step 3: coating the surface of the cell culture plate, the method is the same as in Example 2;

[0101] The above-mentioned high-throughput, in-situ observed cell 3D culture plate is applied to the preparation of liquid marbles, and the preparation of cell culture liquid liquid marbles with small surface tension and large volume includes the following steps:

[0102] Step 1.1: When the volume is large, the low surface tension droplet is easy to adhere to the substrate and cannot form a liquid marble. Therefore, in this example, first use a pipette gun to measure 100 μL of the medium (DMEM+10% FBS+1% double antibody) of MCF7 breast cancer cells, and add it to the ...

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Abstract

The invention belongs to the field of biomedical materials, and particularly relates to preparation and application of a high-throughput in-situ observable cell 3D culture plate. The method comprises the following steps: 1, preparing alkylated SiO2 sol; 2, cleaning the cell culture plate; and 3, coating the alkylated SiO2 sol prepared in the step 1 on the surface of the cell culture plate in the step 2 to obtain the coated cell culture plate. The application is to prepare the single-layer nano-particle structure liquid marbles on the surface of a coated cell culture plate. The prepared super-hydrophobic SiO2 coating has the characteristic of weak binding force, and particles on the outermost layer are easy to transfer. The in-situ preparation of the liquid marbles in the porous plate can be realized by directly rolling the liquid drops of the cell culture fluid on the coating. The prepared liquid marbles can be directly used for 3D culture and high-throughput detection of cells. Compared with a powder source liquid marble, the liquid marble is more transparent, the internal situation can be observed more conveniently, and meanwhile waste and pollution caused by operations such as powder removal and marble transfer are avoided.

Description

technical field [0001] The invention belongs to the field of biomedical materials, and in particular relates to the preparation and application of a high-throughput, in-situ observed cell 3D culture plate. Background technique [0002] Three-dimensional (3D) cell culture plays an important role in basic research and drug experimentation. A large number of studies have shown that three-dimensional cell culture can provide cell growth with living conditions closer to the living body, affect cell migration, aggregation, proliferation and differentiation, and maintain good biochemical and physiological responses of cells. [0003] Compared with the traditional two-dimensional cell culture, the cells in the three-dimensional environment divide in all directions, and the responses of the cells to endogenous and exogenous stimuli, such as temperature, pH, nutrient concentration, oxygen content, etc., are consistent with their in vivo . [0004] Traditional two-dimensional cell cu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12M3/00
CPCC12M25/16C12M25/06C12M23/20
Inventor 李晓光蒋浩浩
Owner NORTHWESTERN POLYTECHNICAL UNIV
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