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Preparation method of centrifugal hydrogel liquid drops

A hydrogel and centrifugal technology, applied in biochemical equipment and methods, tissue culture, microorganisms, etc., can solve the problem of flux, cost, simplicity, lack of uniformity and safety of hydrogel droplets and other issues, to achieve high-throughput repeatability, easy industrial mass production customization, and reduce consumption

Pending Publication Date: 2022-07-05
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] To sum up, the current commonly used preparation technology is related to microfluidics, which is slightly deficient in terms of throughput, cost, and simplicity.
In addition, the hydrogel prepared by the two-phase mixing method will use emulsifiers and cross-linking agents with toxic and side effects, and the uniformity and safety of the prepared hydrogel droplets are not fully guaranteed.

Method used

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  • Preparation method of centrifugal hydrogel liquid drops
  • Preparation method of centrifugal hydrogel liquid drops
  • Preparation method of centrifugal hydrogel liquid drops

Examples

Experimental program
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Effect test

Embodiment 1

[0031] Example 1 Preparation of hydrogel droplets

[0032] A method for preparing centrifugal hydrogel droplets, comprising the steps of:

[0033] 1) Preparation of hydrogel and oil phase

[0034] One or two or more biomacromolecular compounds are selected from GelMA, sodium alginate, and agarose to be compounded into a hydrogel of corresponding mass fraction (or viscosity);

[0035] Mix 1-20 parts: 99-80 parts of EM 180 emulsifier and isopropyl palmitate respectively by volume to obtain an oil phase with certain surface tension and viscosity.

[0036] 2) Preparation of hydrogel droplets

[0037] figure 1 As shown, use PDMS 3 to place capillary 5 (greater than 30 μm in diameter) in a 200 μL pipette tip 4, and fix the pipette tip with capillary in the center of a 15 mL centrifuge tube by snap 2. The centrifuge tube is filled with 5~10 mL of oil phase 6 prepared in step 1), and is used as a container for collecting hydrogel droplets to prevent droplets from being thrown to t...

Embodiment 2

[0038] Example 2 Preparation of cell-loaded hydrogel droplets

[0039] The alternative cells cultured in 2D mode were collected by centrifugation and enzymatically digested to obtain monodisperse cells ( Figure 3.1-3.3 , Figure 4.1-4.3 ) to prepare different numbers of cell solutions. Take the hydrogel prepared in step 1) in Example 1, add 10-1000 cells per 1.0 mL of hydrogel, and add 50-500 μL of sample to the pipette tip of the centrifugal device after mixing. The cells were incubated at 37 °C for 1–5 min, and centrifuged at 100–3000 RCF for 1–5 min to produce a hydrogel solution loaded with cells, and the hydrogel droplets containing cells could be identified under a fluorescence optical microscope ( Figure 3.4 and Figure 4.4 ). Encapsulation of cells in hydrogel droplets can be better illustrated using GFP-bearing cells.

[0040] The above-mentioned centrifugal method for preparing hydrogel droplets loaded with living cells is simple and easy to implement, does no...

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Abstract

The invention discloses a preparation method of centrifugal hydrogel liquid drops, which comprises the following steps: 1) preparing hydrogel, namely selecting one or more biomacromolecular substances to prepare the hydrogel with viscosity; and (2) preparing hydrogel liquid drops: in an oil phase system consisting of isopropyl palmitate containing 1-20% (v / v) EM 180, carrying out centrifugal shearing on the hydrogel prepared in the step (2) to generate the hydrogel liquid drops. According to the other implementation mode, free cells are mixed and digested in the hydrogel in the step 2), and hydrogel liquid drops loaded with the cells are generated through centrifugal shearing. The method provided by the invention can be used for cell 3D culture, has the advantages of low cost, simple process, high throughput, high repeatability and the like compared with the traditional technology, and provides a new preparation method for tissue engineering, cell 3D printing, tumor research, drug development and the like.

Description

technical field [0001] The invention relates to a preparation method of centrifugal hydrogel droplets, belonging to the cross field of biomedical engineering, synthetic biology and microfluidic technology. Background technique [0002] Traditional static cell cultures are often carried out in two-dimensional dishes and contact the surface of glass or plastic dishes. In this way, the expression of genes in the cells is affected and the cells cannot continue to grow and differentiate. In contrast, 3D cell culture can often maintain the differentiation characteristics of cells, and has broad application prospects in tissue chips, organoid models, tumor research and drug development. The use of hydrogel droplets to load cells is an important technical means for three-dimensional cell culture. The production cost, convenience, and reproducibility of cell-loaded hydrogel droplets are directly related to the cost and efficiency of using this technology. [0003] At present, micr...

Claims

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Application Information

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IPC IPC(8): C08J3/00C08J3/12C08J3/075C08L89/00C08L5/04C08L5/12C12N5/00
CPCC08J3/00C08J3/12C08J3/075C12N5/0062C08J2389/00C08J2305/04C08J2305/12C12N2533/54C12N2533/74C12N2533/76
Inventor 唐伟敏何赛灵程潇羽吴咏芳
Owner ZHEJIANG UNIV
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