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A kind of microcarrier for cell culture and its preparation method and application

A cell culture and microcarrier technology, applied in the field of bioengineering, can solve the problems of high polypeptide price, limited wide application, and increased product cost.

Active Publication Date: 2017-11-28
INST OF PROCESS ENG CHINESE ACAD OF SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

In fact, the interaction between cells and microcarriers mainly occurs on the surface of microcarriers, and the protein / polypeptide dispersed inside the microspheres does not play a big role, especially some polypeptides are expensive, and a large amount enters the interior of the microspheres without Exercising the function will cause huge waste, and the product cost will increase accordingly, which limits the wide application of charge-protein / polypeptide dual-action microcarriers

Method used

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  • A kind of microcarrier for cell culture and its preparation method and application
  • A kind of microcarrier for cell culture and its preparation method and application
  • A kind of microcarrier for cell culture and its preparation method and application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0070] This embodiment is the coupling of matrix microspheres and positive charges, and the steps are as follows:

[0071] Get 200g of cross-linked konjac glucomannan microspheres (konjac glucomannan microspheres are prepared according to Chinese invention patent CN101113180 B), add 400mL of NaOH solution with a concentration of 1.2mol / L, and 200mL of epichlorohydrin, at 35°C Constant temperature reaction 2h. After the reaction is completed, wash off the unreacted epichlorohydrin with deionized water, drain it and put it into Na 2 CO 3 -Na 2 HCO 3 Add 10.0 g of diethylamine to the buffer solution, and react at a constant temperature of 35° C. for 18 hours. After the reaction, unreacted substances are washed away with deionized water to obtain aminated (positively charged) konjac glucomannan microspheres.

Embodiment 2

[0073] This embodiment is the coupling of matrix microspheres and positive charges, and the steps are as follows:

[0074] Take 200 g of cross-linked dextran microspheres, add 300 mL of NaOH solution with a concentration of 1.5 mol / L, add 20.0 g of trimethylethanolamine, and react at a constant temperature of 35 ° C for 18 h. After the reaction, unreacted substances are washed away with deionized water to obtain aminated cross-linked dextran microspheres.

Embodiment 3

[0076] This embodiment is the coupling of matrix microspheres and positive charges, and the steps are as follows:

[0077] Take 200 g of agarose microspheres, add 400 mL of NaOH solution with a concentration of 1.2 mol / L, add 12.0 g of 2-diethylaminochloroethane hydrochloride, and react at a constant temperature of 35 ° C for 18 h. After the reaction, unreacted substances are washed away with deionized water to obtain aminated agarose microspheres.

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Abstract

The invention relates to a microcarrier for cell culture and its preparation method and application. The microcarrier has a positive charge and a protein / polypeptide coating. The preparation method of the microcarrier comprises: (1) chemically crosslinking the matrix of the microcarrier with a positive charge modifier; (2) crosslinking the positively charged matrix microsphere obtained in step (1) with protein / polypeptide. The invention also provides the application of the microcarrier in cell culture. The microcarrier of the present invention can limit the thickness of the protein / polypeptide coating, so that the microcarrier can give full play to the role of the protein / polypeptide without completely covering the charges; Protein / polypeptide promotes and regulates the function of cell growth, so as to play the dual role of charge and protein / peptide and effectively regulate its dosage, which has obvious advantages in cell culture and tissue engineering.

Description

technical field [0001] The invention relates to the field of bioengineering, in particular to a microcarrier for cell culture and its preparation method and application. Background technique [0002] Animal cells are the main production vehicle for protein drugs such as vaccines. In the production process of vaccines, the cultivation and expression of animal cells determine the yield and production efficiency of vaccines. In the large-scale culture technology of animal cells, the microcarrier culture method has many advantages: its specific surface area is large, and the cell yield per unit volume of culture solution is high; it adopts uniform suspension culture without nutrient or product gradient; it is easy to observe microcarriers The growth on the surface; the cell harvesting process is relatively simple, the labor intensity is small; the medium utilization rate is high, the floor space is small, and it is easy to scale up. It is currently recognized as the most promis...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/02C12N5/07C12N5/071C12N5/09C12N5/10C08J3/24C08L89/00
Inventor 马光辉周炜清李娟苏志国孙李靖汪少久
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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