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Kit and method used for obtaining full liver stent

A kit and liver technology, applied in medical science, tissue regeneration, prosthesis, etc., can solve the problems of incomplete effect and complex method and process, and achieve the effect of strong buffering ability and stable pH environment.

Inactive Publication Date: 2009-10-28
ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The process of this method is more complicated, and the decellularization effect of a single detergent is not complete enough

Method used

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  • Kit and method used for obtaining full liver stent
  • Kit and method used for obtaining full liver stent
  • Kit and method used for obtaining full liver stent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0026] Example 1. Rat liver decellularization

[0027] 1. Prepare perfusate I and perfusate II:

[0028] Perfusate I: Take 10ml of Triton X-100, dissolve it in 800ml of Tris-HCl with pH 7.5 and 50mM, and dilute to 1000ml to obtain 1% Triton X-100;

[0029] Perfusate II: 10 g of SDS was dissolved in 800 ml of Tris-HCl of pH 7.5 and 50 mM, and the volume was adjusted to 1000 ml to obtain 1% SDS.

[0030] 2. Perfusion to achieve decellularization:

[0031] Insert a cannula from the portal vein of the isolated liver, and perfuse the isolated liver with 1% TritonX-100 perfusion solution at a rate of 10ml / min for 1 hour. At this time, the central area of ​​the liver becomes milky white, and the outflowing perfusate is turbid brown yellow; then Perfuse with double distilled water for 1 hour to fully elute Triton X-100, then perfuse with 1% SDS perfusion solution at a rate of 10ml / min for 3 hours, at this time the liver becomes transparent, and the vascular structure in it can be ...

Embodiment 2

[0032] Example 2. Pig liver perfusion decellularization

[0033] 1. Prepare perfusate I and perfusate II:

[0034] Perfusate I: Dissolve 100ml of Triton X-100 in 800ml of Tris-HCl with pH 8 and 50mM, and dilute to 1000ml to obtain 10% Triton X-100;

[0035] Perfusate II: Dissolve 100 g of SDS in 800 ml of Tris-HCl at pH 8 and 50 mM, and dilute to 1000 ml to obtain 10% SDS.

[0036] 2. Perfusion to achieve decellularization:

[0037] After the portal vein and the hepatic artery were respectively intubated, the above-mentioned perfusate I and perfusate II were perfused simultaneously until the liver became transparent. The perfusion rate used was 50 ml / min. After perfusion, perfuse with double distilled water to fully elute the detergent.

Embodiment 3

[0038] Example 3. Mouse liver decellularization

[0039] 1. Prepare perfusate I and perfusate II:

[0040] Perfusate I: Dissolve 1ml of Triton X-100 in 800ml of Tris-HCl with pH 7.5 and 50mM, and dilute to 1000ml to obtain 0.1% Triton X-100;

[0041] Perfusion solution II: 1 g of ursodeoxycholic acid was dissolved in 800 ml of Tris-HCl of pH 7.5 and 50 mM, and the volume was adjusted to 1000 ml to obtain 0.1% ursodeoxycholic acid.

[0042] 2. Perfusion to achieve decellularization:

[0043]After passing through the superior and inferior hepatic vena cava, the inferior hepatic inferior vena cava was ligated, and the portal vein was cut to perfuse at a rate of 4ml / min. Perfuse according to the method in Example 1.

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Abstract

The invention provides a kit used for obtaining a full liver stent, comprising perfusion liquid I: 0.1 to 10 percent (volume fraction) of Tris-HCl solution of non-ionic detergent; and perfusion liquid II: 0.1 to 10 percent (mass fraction) of Tris-HCl solution of ionic detergent. The invention also provides a method for obtaining the full liver stent by using the kit. The invention leads the substantive organ liver to be acellular to completely and simultaneously reserves the structure and large part of components of extracellular matrix, thus being used as materials of biological stents and being applied to research of biological artificial liver.

Description

technical field [0001] The invention relates to a kit used for obtaining a biological scaffold, in particular to a kit for obtaining a whole liver scaffold. The invention also relates to a method for obtaining a whole liver scaffold using the kit. Background technique [0002] Currently, the only effective treatment for end-stage liver disease (cirrhosis, hepatic encephalopathy, liver cancer) is liver transplantation. Although other methods such as dialysis can prolong human life, they cannot replace all the functions of the liver, and it is difficult to prevent the disease from getting worse. Although the operation of liver transplantation is quite mature at present, there are problems such as the shortage of donor livers and the need for long-term use of immunosuppressants after surgery, which severely limits the clinical application of liver transplantation. Therefore, it is of great practical significance to alleviate the current situation of donor shortage through liv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/56
CPCA61L2430/40A61L27/56A61L27/3839A61L27/3687A61L27/3604
Inventor 康玉占汪艳高毅周焕城
Owner ZHUJIANG HOSPITAL SOUTHERN MEDICAL UNIV
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