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Culture method of artificial liver repair material based on adult stem cell-regenerated silk fibroin tissue engineering scaffold

A technology for regenerating silk fibroin and tissue engineering scaffolds, applied in the field of artificial liver, can solve the problems of horizontal comparison and systematic research of in vitro hepatocyte-like cells, and achieve excellent mechanical properties, high proportion of cells into hepatocytes, and good induction effect. Effect

Active Publication Date: 2019-05-24
DONGHUA UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, for many stem cells derived from adult tissues, no one has done a horizontal comparison and systematic study of their ability to form hepatic-like cells in vitro

Method used

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  • Culture method of artificial liver repair material based on adult stem cell-regenerated silk fibroin tissue engineering scaffold
  • Culture method of artificial liver repair material based on adult stem cell-regenerated silk fibroin tissue engineering scaffold

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Embodiment 1

[0041] A method for cultivating an artificial liver repair material based on an adult stem cell-regenerated silk fibroin tissue engineering scaffold, first, preparing an adult stem cell-regenerated silk fibroin tissue engineering scaffold for a bioartificial liver, the steps are as follows:

[0042] 1) Degumming silkworm cocoons with 0.3% aqueous sodium carbonate solution by mass percentage, dissolving them in a lithium bromide aqueous solution with a molar concentration of 9.0mol / L to obtain regenerated silk fibroin-lithium bromide aqueous solution, centrifuging, filtering, dialysis and Concentrating to obtain a 30% regenerated silk fibroin aqueous solution by mass percentage;

[0043] 2) Using water as a solvent, prepare an aqueous solution of HGF, FGF4 factors and bovine serum albumin, wherein the mass concentrations of HGF and FGF4 factors are both 15 μg / mL, and the mass concentration of bovine serum albumin is 0.6 mg / mL;

[0044] 3) mixing the regenerated silk fibroin aqu...

Embodiment 2

[0065] A method for cultivating an artificial liver repair material based on an adult stem cell-regenerated silk fibroin tissue engineering scaffold, first, preparing an adult stem cell-regenerated silk fibroin tissue engineering scaffold for a bioartificial liver, the steps are as follows:

[0066] 1) Degumming the tussah silkworm cocoons with 0.4% aqueous sodium carbonate solution by mass percentage, dissolving them in a lithium bromide aqueous solution with a molar concentration of 9.1mol / L to obtain a regenerated silk fibroin-lithium bromide aqueous solution, centrifuging, filtering, dialysis and Concentrating to obtain a 32% regenerated silk fibroin aqueous solution by mass percentage;

[0067] 2) Using water as a solvent, prepare an aqueous solution of HGF, FGF4 factors and bovine serum albumin, wherein the mass concentrations of HGF and FGF4 factors are both 16 μg / mL, and the mass concentration of bovine serum albumin is 0.7 mg / mL;

[0068] 3) mixing the regenerated sil...

Embodiment 3

[0077] A method for cultivating an artificial liver repair material based on an adult stem cell-regenerated silk fibroin tissue engineering scaffold, first, preparing an adult stem cell-regenerated silk fibroin tissue engineering scaffold for a bioartificial liver, the steps are as follows:

[0078] 1) Degumming castor silkworm cocoons with 0.5% aqueous sodium carbonate solution by mass percentage, dissolving them in lithium bromide aqueous solution with a molar concentration of 9.2mol / L to obtain regenerated silk fibroin-lithium bromide aqueous solution, centrifuging, filtering, and dialyzing the solution and concentrated to obtain a 34% regenerated silk fibroin aqueous solution by mass percent;

[0079] 2) Using water as a solvent, prepare an aqueous solution of HGF, FGF4 factors and bovine serum albumin, wherein the mass concentrations of HGF and FGF4 factors are both 18 μg / mL, and the mass concentration of bovine serum albumin is 0.8 mg / mL;

[0080] 3) mixing the regenerat...

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Abstract

The invention relates to a culture method of an artificial liver repair material based on an adult stem cell-regenerated silk fibroin tissue engineering scaffold. The method mainly comprises two steps: first step, the adult stem cell-regenerated silk fibroin tissue engineering scaffold which is mixed with HGFs (hepatocyte growth factors), FGF4 (fibroblast growth factors 4), bovine serum albumin and a regenerated silk fibroin material and applied to a bioartificial liver is prepared with an electrostatic spinning method; second step, the artificial liver repair material is cultured on the basis of the adult stem cell-regenerated silk fibroin tissue engineering scaffold, and ADSCs (adipose tissue-derived stromal cells) or BMSCs (bone mesenchymal stem cells) adhere to and proliferate on the regenerated silk fibroin material and are induced to form liver-like cells under the action of scaffold sustained-release HGFs and FGF4. The prepared artificial liver repair material based on the adult stem cell-regenerated silk fibroin tissue engineering scaffold has good biocompatibility, excellent biodegradability and excellent mechanical property and is a good alternative scaffold material for the bioartificial liver.

Description

technical field [0001] The invention belongs to the technical field of artificial liver, and relates to a method for cultivating an artificial liver repair material based on an adult stem cell-regenerated silk fibroin tissue engineering scaffold. Background technique [0002] my country is a big country with hepatitis B, and the carrier rate of hepatitis B virus is as high as nearly 10%. With the development of economy and the change of lifestyle, alcoholic liver disease caused by heavy drinking is also on the rise year by year; Drug abuse and drug-induced liver disease are also increasing year by year. These huge numbers of liver disease patients, after a certain course of disease, some patients will progress to end-stage liver disease or acute liver failure. Orthotopic liver transplantation is the only effective treatment for this kind of disease, but due to many factors such as lack of donor liver, immune rejection, surgical risk, and high cost, its wide application in cl...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/38A61L27/22
CPCA61L27/22A61L27/3834A61L2300/252A61L2430/28
Inventor 张耀鹏阎丽徐丽娟黄利邵惠丽胡学超
Owner DONGHUA UNIV
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