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Macroporous microcarrier for cell cultivation, preparation method and usage thereof

A technology of microcarriers and microspheres, applied in biochemical equipment and methods, tissue culture, microorganisms, etc., can solve the problems that cells are susceptible to mechanical damage, unevenness, and cells are easy to fall off and adhere to the wall, and achieve long-term maintenance of cell activity and Function, good slow degradation, good mechanical properties

Active Publication Date: 2010-01-13
ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the existing microcarriers cannot maintain cell growth for a long time, and the cells are easily damaged by machinery. When the cells are enlarged and cultured, they are easy to fall off and adhere unevenly. Therefore, it is necessary to provide a new type of microcarrier to overcome the defects in the prior art.

Method used

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  • Macroporous microcarrier for cell cultivation, preparation method and usage thereof
  • Macroporous microcarrier for cell cultivation, preparation method and usage thereof
  • Macroporous microcarrier for cell cultivation, preparation method and usage thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1) Preparation of silk fibroin solution: boil 75g of raw silk in 4L of 5g / L sodium carbonate solution for 1 hour, then wash with a large amount of deionized water to remove sericin, and dry at 60-70°C to obtain silk fibroin. Dissolve an appropriate amount of silk fibroin in a ternary solution of calcium chloride / water / ethanol (molar ratio 1:8:2) at 80±2°C, and dialyze with deionized water at room temperature for 3 days to remove salt and ethanol in the solution , filter to remove insoluble impurities, and prepare an aqueous solution of silk fibroin. Stir and concentrate the silk fibroin aqueous solution at 50±2°C and 60 rpm to obtain a silk fibroin solution with a concentration of about 7-10w / v%;

[0031] 2) Preparation of silk fibroin-chitosan mixed solution: prepare 4w / v% chitosan acetic acid aqueous solution, prepare silk fibroin-chitosan mixed solution, silk fibroin, chitosan mass ratio is 7: 3, final The concentration is 5w / v%;

[0032] 3) Take a certain amount ...

Embodiment 2

[0039] 1) Preparation of silk fibroin solution: boil 75g of raw silk in 4L of 5g / L sodium carbonate solution for 1 hour, then wash with a large amount of deionized water to remove sericin, and dry at 60-70°C to obtain silk fibroin. Dissolve an appropriate amount of silk fibroin in a ternary solution of calcium chloride / water / ethanol (molar ratio 1:8:2) at 80±2°C, and dialyze with deionized water at room temperature for 3 days to remove salt and ethanol in the solution , filter to remove insoluble impurities, and prepare an aqueous solution of silk fibroin. Stir and concentrate the silk fibroin aqueous solution at 50±2°C and 50-60 rpm to obtain a silk fibroin solution with a concentration of about 7-10w / v%;

[0040]2) Preparation of silk fibroin-chitosan mixed solution: prepare 4w / v% chitosan acetic acid aqueous solution, prepare silk fibroin-chitosan mixed solution, silk fibroin, chitosan mass ratio is 6: 4, final The concentration is 4w / v%;

[0041] 3) Take a certain amoun...

Embodiment 3

[0048] 1) Preparation of silk fibroin solution: boil 75g of raw silk in 4L of 5g / L sodium carbonate solution for 1 hour, then wash with a large amount of deionized water to remove sericin, and dry at 60-70°C to obtain silk fibroin. Dissolve an appropriate amount of silk fibroin in a ternary solution of calcium chloride / water / ethanol (molar ratio 1:8:2) at 80±2°C, and dialyze with deionized water at room temperature for 3 days to remove salt and ethanol in the solution , filter to remove insoluble impurities, and prepare an aqueous solution of silk fibroin. Stir and concentrate the silk fibroin aqueous solution at 50±2°C and 60 rpm to obtain a silk fibroin solution with a concentration of about 7-10w / v%;

[0049] 2) Preparation of silk fibroin-chitosan mixed solution: prepare 4w / v% chitosan acetic acid aqueous solution, prepare silk fibroin-chitosan mixed solution, silk fibroin, chitosan mass ratio is 5: 5, final The concentration is 4w / v%;

[0050] 3) Take a certain amount ...

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Abstract

The invention provides fibroin / chitosan macroporous microcarrier prepared by taking silk fibroin as main raw material, a preparation method and the usage thereof. The macroporous microcarrier takes on sphere, has the diameter of 200-500mu m and the aperture of 40-80mu m, and made of the silk fibroin and the chitosan under the action of crosslinking agent. The fibroin / chitosan macroporous microcarrier takes the fibroin as main ingredient, and the silk fibroin has good biocompatibility, slow degradation and excellent mechanical property in a wet state, so that the structure of the microcarrier can be maintained by the silk fibroin for a longer time; therefore, the silk fibroin not only can support the growth of a large number of cells, but also can maintain the activity and the functions of the cells for a long time, thus having unique function in the high density cultivation of animal cells for a long time.

Description

technical field [0001] The present invention relates to a macroporous microcarrier for large-scale culture of animal cells and its preparation method and application, in particular to a silk fibroin / chitosan macroporous microcarrier prepared from silk fibroin as the main raw material and its preparation Methods and uses. Background technique [0002] Microcarrier culture is currently recognized as the most promising large-scale culture technology for animal cells. It has both the advantages of suspension culture and adherent culture, and is easy to scale up. Macroporous microcarriers immobilize cells to grow in the holes, which can maximize the specific surface area. Compared with other methods, its specific surface area is larger, which is several times or even dozens of times that of solid microcarriers; cells grow in the holes , is protected, and the shear damage is small; the three-dimensional growth of cells will further increase the cell density, and the cell density ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C08L89/00C08L5/08C08J9/28C08J3/24C12N5/08C12N11/00
Inventor 龚独辉潘明新高毅周焕城蒋泽生
Owner ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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