Porous chitosan microcarrier as well as preparation method and application thereof

A technology of porous chitosan and microcarriers, applied in the direction of microcarriers, general culture methods, biochemical equipment and methods, etc., can solve the problem of inability to form cell-cell connections, inability to support a large number of cell growth, unfavorable cell adhesion and growth To achieve good biocompatibility, uniform size, maintain vitality and function

Active Publication Date: 2018-05-18
EZHOU INST OF IND TECH HUAZHONG UNIV OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The present invention overcomes the low porosity of porous chitosan microcarriers in the prior art, the lack of interconnected spaces on the surface and inside, which is not conducive to cell adhesion and growth, cannot support a large number of cell growth, and cannot form cell-cell connections. Can not realize true three-dimensional culture and other defects, provide a porous chitosan microcarrier and its preparation method and application

Method used

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  • Porous chitosan microcarrier as well as preparation method and application thereof
  • Porous chitosan microcarrier as well as preparation method and application thereof
  • Porous chitosan microcarrier as well as preparation method and application thereof

Examples

Experimental program
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Embodiment 1

[0046] The preparation of embodiment 1 porous chitosan microcarrier

[0047] 1, prepare 1% (w / v) chitosan deacetylation degree 95%, viscosity-average molecular weight 100,000 (supplier is ) of acetic acid aqueous solution as the dispersed phase, wherein the acetic acid content is 1% (w / v); the petroleum ether solution of the preparation 5% (w / v) emulsifier is used as the continuous phase, wherein the emulsifier is composed of Span 80 and Tween 60 according to 20:1 (w / w) mixed composition; prepare 1% (w / v) ethanol / water solution of sodium hydroxide as a phase transition solution, wherein the ratio of ethanol to water is 14:1 (v / v).

[0048] 2. Slowly add 10 mL of the above dispersed phase solution into 50 mL of the continuous phase solution, and emulsify at 40°C for 4 hours under stirring at 1000 r / min to obtain a W / O emulsion.

[0049] 3. The resulting W / O emulsion was quenched at -20°C for 4 hours to conduct thermally induced phase separation.

[0050] 4. Pour 100mL of the...

Embodiment 2

[0052] The preparation of embodiment 2 porous chitosan microcarriers

[0053] 1, the acetic acid aqueous solution of preparation 1% (w / v) chitosan (deacetylation degree 95%, viscosity-average molecular weight 100,000) is as dispersed phase, wherein acetic acid content 1% (w / v); Preparation 7% (w / v) / v) Petroleum ether solution of emulsifier as continuous phase, wherein emulsifier is composed of Span 80 and Tween 60 mixed in 24:1 (w / w); prepare 1.5% (w / v) ethanol / water solution of sodium hydroxide as Phase change liquid, wherein the ratio of ethanol to water is 20:1 (v / v).

[0054] 2. Slowly add 10 mL of the above dispersed phase solution into 30 mL of the continuous phase solution, and emulsify at 30°C for 4 hours under stirring at 1000 r / min to obtain a W / O emulsion.

[0055] 3. The resulting W / O emulsion was quenched at -50°C for 2 hours to conduct thermally induced phase separation.

[0056] 4. Pour 80mL of the above-mentioned phase transition liquid into the above-mentio...

Embodiment 3

[0058] The preparation of embodiment 3 porous chitosan microcarriers

[0059] 1, prepare 3% (w / v) chitosan (deacetylation degree is greater than 80%, viscosity-average molecular weight 150,000, supplier is ) of acetic acid aqueous solution as the dispersed phase, wherein the acetic acid content is 3% (w / v); the petroleum ether solution of the preparation 7% (w / v) emulsifier is used as the continuous phase, wherein the emulsifier is composed of Span 80 and Tween 60 according to 24:5 (w / w) mixed composition; 3% (w / v) sodium hydroxide ethanol / water solution was prepared as a phase transition solution, wherein the ratio of ethanol to water was 20:5 (v / v).

[0060] 2. Slowly add 10 mL of the above dispersed phase solution into 100 mL of the continuous phase solution, and emulsify at 50°C and 1500 r / min for 2 hours to obtain a W / O emulsion.

[0061] 3. The resulting W / O emulsion was quenched at -5°C for 6 hours to conduct thermally induced phase separation.

[0062] 4. Pour 110mL...

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Abstract

The invention discloses a porous chitosan microcarrier as well as a preparation method and an application thereof. The porous chitosan microcarrier takes chitosan as a main component, the diameter ofthe porous chitosan microcarrier is 100-300 mu m, interconnected pores are formed in the surface and the interior of the porous chitosan microcarrier, the diameter of the pores is 10-70 mu m, and theporosity is 95% or higher. The porous chitosan microcarrier is uniform in size, the interconnected pores are formed in the surface and the interior, and the highest porosity can reach 95% or higher. With adoption of the porous chitosan microcarrier for culturing cells, the cells can not only be adhered to the surface of the carrier, but also can enter the internal pores to grow, effective cell-cell connection is formed in three-dimensional space, real three-dimensional culture is realized, in vivo environment is effectively simulated, vitality and function of cells are better maintained, and the microcarrier has good biocompatibility.

Description

technical field [0001] The invention belongs to the field of natural polymer porous materials, and in particular relates to a porous chitosan microcarrier and its preparation method and application. Background technique [0002] Microcarrier technology is an effective means to achieve large-scale cell culture. At present, some solid or porous microcarriers have been commercialized abroad, such as Cytodex, Cutispher, etc. At the same time, there are many kinds of microcarriers in the laboratory research stage. Compared with traditional plate culture, these microcarriers can provide sufficient adhesion sites for adherent cells and realize large-scale suspension culture of cells. Three-dimensional space forms effective cell-cell connections, thus failing to achieve true three-dimensional cell culture. [0003] Chitosan is a deacetylated product of chitin, which exists widely in nature. It has good biocompatibility, blood compatibility and microbial degradability, and has broad...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08J9/24C08L5/08C12N5/02C12N5/071
CPCC08J9/24C08J2305/08C12N5/0075C12N5/0671C12N2531/00
Inventor 杨光黄丽霞肖林郑瑞珠刘海清
Owner EZHOU INST OF IND TECH HUAZHONG UNIV OF SCI & TECH
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