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Preparation method of single-cell microgel

A microgel, single-cell technology, applied in the direction of immobilization on/in an organic carrier, can solve the problems of high manufacturing cost and use cost

Active Publication Date: 2021-10-22
SHENZHEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] At present, single-cell microgels are mainly prepared by microfluidic technology. Although microfluidic technology can achieve precise single-cell encapsulation, the microfluidic chip used in microfluidic technology is composed of precise microvalve, Micropumps and micropipes of tens of microns are processed and assembled, and each microfluidic chip is only suitable for a small number of specific cells and specific occasions, and the manufacturing cost and use cost are high

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Examples

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preparation example Construction

[0062] The preparation method of the above-mentioned single-cell microgel has at least the following advantages:

[0063] (1) Low cost: the raw materials are at least one of calcium carbonate nanoparticles and barium carbonate nanoparticles, alginate and polycation compounds, the raw materials are easy to obtain and cheap, and the preparation method of the above-mentioned single-cell microgel Do not rely on expensive microfluidic chips, low cost.

[0064] (2) Easy to operate: The above-mentioned single-cell microgel preparation method can realize the preparation of single-cell microgel by solid-liquid mixing and solid-liquid separation, and the operation is simple; and, the traditional single-cell microgel prepared by microfluidic chip Microgels are not easy to collect, separate and process. However, when the single-cell microgels are prepared by the above-mentioned single-cell microgel preparation method, they can be collected, separated and processed by centrifugation. In a...

Embodiment 1

[0071] (1) Sample and reagent preparation:

[0072] Sodium alginate solution: Mix sodium alginate, sodium alginate marked with FITC (FITC sodium alginate) and water evenly to prepare a sodium alginate solution. In the sodium alginate solution, the mass percentage of sodium alginate is 1% sodium alginate, the mass percentage of FITC-sodium alginate is 0.05%, and the cells are Hela cells;

[0073] Polyethyleneimine dispersion: 0.0008g polyethyleneimine (Aladdin E107079, weight average molecular weight 10000) and 1mL deionized water were dispersed under 80kHz ultrasonic for 3 minutes to prepare polyethyleneimine dispersion; In the solution, the concentration of polyethyleneimine is 0.8mg / mL;

[0074] It is standby to take by weighing 0.0099g median particle diameter to be the calcium carbonate nanoparticle of 20nm;

[0075] Cell suspension: 1mL cell suspension whose cell membrane is labeled with red probe (DID), the cell density is 1×10 6 cells / mL; Electron micrographs of cell...

Embodiment 2

[0085] The preparation method of the single-cell gel in this example is roughly the same as the preparation method of the single-cell gel in Example 1, the difference is that in this example, the concentration of polyethyleneimine in the polyethyleneimine dispersion is 0.6 mg / mL, the potential of positively charged calcium carbonate nanoparticles in water is 14.1mV.

[0086] The single-cell gel prepared according to the preparation method of this example is as follows: Figure 9 to Figure 11 shown. Figure 9 is the image under the red channel; Figure 10 is the image under the green channel; Figure 11 is the image under the merged channel.

[0087] Depend on Figure 9 to Figure 11 It can be seen that the single-cell microgel of this example was successfully prepared, there was gel on the cell surface, and there was fluorescence under the red and green channels. Through the fluorescence of the combined channel, it was found that the encapsulation rate of the single-cell m...

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Abstract

The invention relates to a preparation method of single-cell microgel. The preparation method of the single-cell microgel comprises the following steps: processing a cross-linking agent so that the cross-linking agent can carry positive charges to prepare the cross-linking agent with the positive charges, wherein the cross-linking agent is selected from at least one of calcium carbonate nano-particles and barium carbonate nano-particles; mixing a cross-linking agent with positive charges with cells to prepare cells adsorbed with the cross-linking agent; and mixing and reacting the cells adsorbed with the cross-linking agent with an alginate solution to prepare the single-cell microgel. The preparation method of the single-cell microgel is simple and low in cost.

Description

technical field [0001] The invention relates to the technical field of biological materials, in particular to a preparation method of single-cell microgel. Background technique [0002] Single-cell-laden microgels contain only one cell surrounded by a hydrogel matrix. The gel layer of single-cell microgel has the following characteristics: (1) The gel layer wrapped outside the cells is a biological semipermeable membrane, which can not only realize immune isolation, prevent macromolecular immune substances and immune cell attacks, but also allow Metabolites, small molecule nutrients and cell active substances can freely enter and exit microcapsules, which makes single-cell microgels have great advantages in cell therapy. (2) The gel layer can provide conditions such as the three-dimensional structure required for cell growth and a cell matrix (such as hydrogel) with good biocompatibility. Compared with the traditional two-dimensional monolayer cell culture, three-dimension...

Claims

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Application Information

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IPC IPC(8): C12N11/10C12N11/04
CPCC12N11/10C12N11/04
Inventor 孔湉湉张姗姗刘洲杜伟翔庄艳逢
Owner SHENZHEN UNIV
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