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Preparation method of accellular sheep valved vein stent material

A scaffold material and decellularization technology, which is applied in the field of preparation of decellularized sheep vein scaffold materials with valves, can solve the problems of difficult acquisition of decellularized scaffolds, and achieve the effects of good water solubility, high economic benefits, and strong usability

Inactive Publication Date: 2017-03-08
SHAANXI EYOUNG TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, it is not easy to obtain allogeneic decellularized scaffolds, and researchers are still focusing on xenogeneic scaffolds

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0014] A method for preparing a decellularized sheep vein stent material with a valve, comprising the following steps:

[0015] (1) Dozens of valved femoral veins of normal adult sheep were cut under sterile conditions and trimmed to about 2 cm;

[0016] (2) After cleaning with PBS, place in 0.5% Triton X-100+0.05% NH 4 After shaking in 0H mixed liquid at 4°C for 3 days, shake and rinse in ultrapure water for 3 days at 4°C;

[0017] (3) Then use DNase+RNase to act at 37°C for 12 hours, rinse with ultrapure water for 2 hours, put the obtained acellular matrix material into an Eppendorf tube, seal it with a parafilm, 60 Sterilized by CO irradiation, stored at minus 60-100°C;

[0018] (4) Put the decellularized scaffold material into a 6-well plate, press 6cm 2 / ml, surface area / volume ± 10% ratio, add lymphocyte culture medium;

[0019] (5) Incubate at 37°C for (24±2) h to prepare scaffold material extract, put the prepared scaffold material extract into an Eppendorf tube, s...

Embodiment 2

[0021] A method for preparing a decellularized sheep vein stent material with a valve, comprising the following steps:

[0022] (1) Dozens of valved femoral veins of normal adult sheep were cut under sterile conditions and trimmed to about 2 cm;

[0023] (2) After cleaning with PBS, place in 0.5% Triton X-100+0.05% NH 4 After shaking in 0H mixed liquid at 4°C for 3 days, shake and rinse in ultrapure water for 3 days at 4°C;

[0024] (3) Then use DNase+RNase to act at 37°C for 12 hours, rinse with ultrapure water for 2 hours, put the obtained acellular matrix material into an Eppendorf tube, seal it with a parafilm, 60 Sterilized by CO irradiation, stored at minus 60-100°C;

[0025] (4) Put the decellularized scaffold material into a 6-well plate, press 6cm 2 / ml, surface area / volume ± 10% ratio, add lymphocyte culture medium;

[0026] (5) Incubate at 37°C for (24±2) h to prepare scaffold material extract, put the prepared scaffold material extract into an Eppendorf tube, s...

Embodiment 3

[0029] A method for preparing a decellularized sheep vein stent material with a valve, comprising the following steps:

[0030] (1) Dozens of valved femoral veins of normal adult sheep were cut under sterile conditions and trimmed to about 2 cm;

[0031] (2) After cleaning with PBS, place in 0.5% Triton X-100+0.05% NH 4 After shaking in 0H mixed liquid at 4°C for 3 days, shake and rinse in ultrapure water for 3 days at 4°C;

[0032] (3) Then use DNase+RNase to act at 37°C for 12 hours, rinse with ultrapure water for 2 hours, put the obtained acellular matrix material into an Eppendorf tube, seal it with a parafilm, 60 Sterilized by CO irradiation, stored at minus 60-100°C;

[0033] (4) Put the decellularized scaffold material into a 6-well plate, press 6cm 2 / ml, surface area / volume ± 10% ratio, add lymphocyte culture medium;

[0034] (5) Incubate at 37°C for (24±2) h to prepare scaffold material extract, put the prepared scaffold material extract into an Eppendorf tube, s...

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Abstract

The invention relates to a preparation method of an accellular sheep valved vein stent material. The method is characterized by comprising the following steps: cutting off tens of normal adult sheep valved veins under aseptic conditions, and pruning into 2cm or so; cleaning with a PBS (phosphate buffer solution), putting into a 0.5%Triton X-100+0.05%NH40H mixed liquid, shaking at 4 DEG C for 3 days, putting into ultrapure water, and rinsing by shaking at 4 DEG C for 3 days. The accellular sheep valved vein stent material prepared by the method has the advantages of favorable water solubility, no damage to cells and stable properties in the culture fluid, and can not be spontaneously oxidized or reduced easily. The method has the advantages of simple steps, high usability and high economic benefits.

Description

technical field [0001] The invention relates to a preparation method of a decellularized sheep vein with valve bracket material. Background technique [0002] The rapid development of tissue engineering has made it possible to construct tissue engineered veins with valves. During the construction, the decellularized scaffold is favored by many researchers because of its unparalleled three-dimensional space structure and good cell affinity. At present, it is not easy to obtain allogeneic decellularized scaffolds, and researchers are still focusing on xenogeneic scaffolds. Obviously, immunogenic substances such as residual proteins and cell components during the decellularization process will inevitably cause different degrees of immune responses to the recipients. Therefore, the immunogenicity evaluation of decellularized scaffold materials has become an important link in the safety evaluation of scaffolds. Contents of the invention [0003] The invention provides a meth...

Claims

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Application Information

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IPC IPC(8): A61L27/36
CPCA61L27/3625A61L27/3687
Inventor 张俊
Owner SHAANXI EYOUNG TECH
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