Preparation method of photoresponsive intelligent gel microspheres for three-dimensional cell culture

A three-dimensional culture and smart gel technology, applied in animal cells, chemical instruments and methods, alkali metal oxides/hydroxides, etc., can solve problems such as difficult to study a single process, difficult to study intermediate processes, etc.

Inactive Publication Date: 2014-08-06
SOUTHEAST UNIV
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Problems solved by technology

Ordinary cell culture is often inconsistent with the situation in vivo because cells proliferate and gradually lose their original traits in the changed environment in vitro, while animal experiments are completely carried out in vivo, with various constraints in the body and the interaction between the internal and external environments. Cell culture becomes complicated, single processes difficult to study, and intermediary processes difficult to study

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  • Preparation method of photoresponsive intelligent gel microspheres for three-dimensional cell culture
  • Preparation method of photoresponsive intelligent gel microspheres for three-dimensional cell culture
  • Preparation method of photoresponsive intelligent gel microspheres for three-dimensional cell culture

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preparation example Construction

[0025] Preparation of modified azobenzenes: carboxylation of 4-aminoazobenzene and its polymerization with arginine-glycine-aspartic acid (RGD) (Azo-RGD) (Reference: Z. Zhangetal. Effects of immobilizing sites of RGD peptides in amphiphilic block copolymers on of celladhesion.Biomaterials, 2010, 7873-7882): 1.97g of 4-aminoazobenzene and 1.20g of succinic anhydride were dissolved in 25mL of acetone, 0.97g of anhydrous pyridine was added, the reaction was heated at 60°C for 6h, filtered and vacuum Dry for 24 hours to prepare carboxylated 4-aminoazobenzene Azo-COOH.

[0026] Take 0.137g (0.46mmoL) of the prepared Azo-COOH, dissolve it in 100mL of dichloromethane, and dissolve 0.225g (1.17mmoL) of 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide salt Add salt (EDC) and 0.135g (1.17mmoL) N-hydroxysuccinimide (NHS) to the above solution, stir at room temperature for 24h, remove dichloromethane by rotary evaporation, add 10mL of acetonitrile to dissolve the polymer, and then Add 0.1...

Embodiment 1

[0028] Preparation method of light-responsive smart gel microspheres for three-dimensional cell culture:

[0029] (1) Preparation of aqueous phase solution: Take 0.7800g (5mmoL) poly(methyl vinyl ether-maleic anhydride) and 0.2270g (0.2mmoL) β-cyclodextrin, dissolve in 10mLN, N-dimethyl formaldehyde In amide (DMF), under the protection of nitrogen, heat the reaction at 80°C for 9 hours, and rotate to evaporate to remove the N,N-dimethylformamide solvent; 1.0000g (0.05mmoL) of polyethylene glycol (20000) and 0.0612g (0.2mmoL) of agarose (AG) was dissolved in 10mL of water, added to the above reactants, and shaken until dissolved to prepare an aqueous phase solution with a concentration of 0.2068g / mL for later use.

[0030] (2) Formation of microspheres: Weigh 50ml of simethicone and pour it into a 100ml tall beaker, heat to 80°C, and at a stirring speed of 1000rpm, use a dropper to dissolve 2.5mL of the aqueous phase solution prepared in step (2) Slowly drop into simethicone o...

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Abstract

The invention relates to a preparation method of photoresponsive intelligent gel microspheres for three-dimensional cell culture. The preparation method is characterized by comprising the following steps: dissolving poly(methyl vinyl ether-maleic anhydride) and beta-cyclodextrin into a solvent, namely N,N-dimethylformamide, carrying out heating reaction under the protection of nitrogen, and removing the solvent; adding a mixture water solution of polyethylene glycol 20000 and agarose into the reactant, and vibrating until the reactant is dissolved, so as to prepare a water-phase solution for later use; dropwise adding the water-phase solution into oil-phase simethicone, carrying out heating, stirring, reaction and standing, removing supernate, cleaning microspheres deposited at the bottom of a container, and carrying out vacuum drying, so as to obtain the gel microspheres; putting the gel microspheres into Azo-RGD methane water solution, vibrating for 24 hours under the room temperature, repeatedly rinsing by virtue of deionized water, and drying by virtue of N2. By utilizing the gel microspheres prepared by virtue of the preparation method, growing environments of cells in vivo can be simulated, and the adsorption and controllable separation of the cells on the microspheres can be realized.

Description

technical field [0001] The invention relates to the preparation technology of polymer microspheres, in particular to a method for preparing light-responsive intelligent gel microspheres by a reverse suspension polymerization method, and the obtained microspheres can be used as cell culture microcarriers. Background technique [0002] Three-dimensional cell culture technology is between monolayer cell culture and animal experiments. Compared with traditional two-dimensional cell culture (2D) culture, it is mainly based on the three-dimensional culture model of common scaffolds, which is closer to the microenvironment of cell growth in vivo and can be more efficient. It can well simulate the natural environment of cell growth in vivo, so it is more realistic and reliable in the study of cell growth, differentiation, migration, etc. Ordinary cell culture gradually loses its original characteristics due to the proliferation of cells in a changed environment in vitro, which is of...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/26B01J20/28B01J20/30C12N5/07
Inventor 张天柱马丹丹顾宁
Owner SOUTHEAST UNIV
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