Macroporous micro-carrier with liver cell specificity as well as preparation method and applications thereof

A technology of microcarriers and spheres, applied in the field of hepatocyte-specific macroporous microcarriers and its preparation, can solve the problems of inability to induce and improve the adhesion performance of hepatocytes, easy detachment of cells, and lack of hepatocyte specificity

Active Publication Date: 2011-04-13
ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, in the culture of hepatocytes, the existing macroporous microcarriers are not specific for hepatocytes, cannot induce and improve the adhesion of hepatocytes on the microcarriers, and the cells are easy to fall off when they are enlarged and cultured. Specific macroporous microcarriers are of great significance for the large-scale culture of hepatocytes in vitro

Method used

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  • Macroporous micro-carrier with liver cell specificity as well as preparation method and applications thereof
  • Macroporous micro-carrier with liver cell specificity as well as preparation method and applications thereof
  • Macroporous micro-carrier with liver cell specificity as well as preparation method and applications thereof

Examples

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Embodiment 1

[0033] 1) Preparation of silk fibroin: boil 75g of raw silk in 2L of 5g / L sodium carbonate solution for 0.5 hour, repeat twice, then wash with a large amount of distilled water to remove sericin, dry at 60-70°C, Get 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, dialyze in distilled water for 3 days at room temperature, and remove the 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%;

[0034] 2) Preparation of galactosylated chitosan (GC): Weigh 2.2g of chitosan, dissolve it in 30-40ml 2.0% acetic acid aqueous solution, add an appropriate amount of TEMED / HCl buffer solution (pH 4.7) to dissolve the shell Dilute the polysaccharide solution to 4w / v%, then ...

Embodiment 2

[0042] 1) Preparation of silk fibroin: boil 75g of raw silk in 4L of 5g / L sodium carbonate solution for 0.5 hours, repeat twice, then wash with a large amount of distilled water to remove sericin, dry at 60-70°C, Get 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°C, and dialyze in distilled water for 3 days at room temperature to remove salt, ethanol and other substances in the solution. small molecules, and then filtered to remove insoluble impurities to 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%;

[0043] 2) Preparation of galactosylated chitosan (GC): take 1.1g chitosan, dissolve it in 15-20ml 2.0% acetic acid aqueous solution, and then add an appropriate amount of TEMED / HCl buffer solution (pH4.7) to dissolve Dilute the chito...

Embodiment 3

[0051] 1) Preparation of silk fibroin: boil 75g of raw silk in 2L of 5g / L sodium carbonate solution for 0.5 hour, repeat twice, then wash with a large amount of distilled water to remove sericin, dry at 60-70°C, Get 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, dialyze in distilled water for 3 days at room temperature, and remove the 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%;

[0052] 2) Preparation of galactosylated chitosan (GC): take 2.2g chitosan, dissolve it in 30-40ml 2.0% acetic acid aqueous solution, and then add an appropriate amount of TEMED / HCl buffer solution (pH4.7) to dissolve Dilute the chitosan solution to 4w / v%, then add 0.14g NH...

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Abstract

The invention provides a macroporous micro-carrier with the liver cell specificity mainly made from SF/GC (Silk Fibroin/Galactosyl Chitosan) as well as a preparation method and the applications thereof. The macroporous micro-carrier is a sphere prepared from SF and GC under the action of a crosslinker, wherein the content of SF is 50-80 percent by weight of the sphere, and the content of GC is 15-40 percent by weight of the sphere, and the micro-carrier has the diameter of 200-500 microns and the aperture of 40-80 microns. The micro-carrier induces and improves the adhesion performance of the SF/GC on a macroporous micro-carrier support material to have the liver cell specificity and is suitable for culturing mass liver cells.

Description

technical field [0001] The present invention relates to a macroporous microcarrier for cell culture, its preparation method and application, in particular to a liver cell-specific macroporous microcarrier with silk fibroin and galactosylated chitosan as main raw materials and its preparation Methods and uses. Background technique [0002] The bioartificial liver is an extracorporeal artificial liver support system developed in the late 1980s. The core part of the bioreactor is a combination of liver cells and biosynthetic materials. How to establish a bioreactor culture system with high quality, sufficient quantity, and strong safety is the most important and most difficult core technical issue in the field of bioartificial liver research. [0003] Hepatocytes are polar, anchorage-dependent cells that require an insoluble extracellular matrix to survive, reorganize, proliferate, and function. Hepatocytes in vivo are in a three-dimensional environment, and the interaction b...

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张志胡志伟
Owner ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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