Preparation method of decellularized biological omentum and decellularized biological omentum prepared therefrom

A biological omentum and decellularization technology, which is applied in the field of tissue engineering and biomedical materials, can solve the problems of high cost and cumbersome method steps, and achieve the effects of mild method conditions, improved decellularization efficiency, and simple operation

Active Publication Date: 2017-10-27
北京奥泰康医药技术开发有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Because the structure and function of omentum tissue are quite different from skin, blood vessels, valves and other tissues, traditional decellularization methods, such as repeated freezing and thawing, surfactant method, enzyme digestion method, etc., cannot be fully applied to the omentum Decellularization of tissues
And the method steps are cumbersome, the cost is high, and it is not suitable for batch production

Method used

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  • Preparation method of decellularized biological omentum and decellularized biological omentum prepared therefrom
  • Preparation method of decellularized biological omentum and decellularized biological omentum prepared therefrom
  • Preparation method of decellularized biological omentum and decellularized biological omentum prepared therefrom

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preparation example Construction

[0032] According to one aspect of the present invention, a method for preparing decellularized biological omentum is provided, which includes the step of soaking the greater omentum with a buffer solution containing sodium dodecyl sulfate and DNase for decellularization.

[0033] In some embodiments, shaking is also performed during the decellularization step.

[0034] In some embodiments, a degreasing step is also included. In an exemplary embodiment, the step of defatting treatment is performed prior to the step of decellularization treatment. In some preferred embodiments, the step of degreasing treatment is carried out using a mixed solution of methanol, chloroform and ether at a volume ratio of 1:1:0.2-1. In certain more preferred embodiments, the volume ratio of methanol, chloroform and diethyl ether is 1:1:0.4-0.7. In the most preferred embodiment, the volume ratio of methanol, chloroform and diethyl ether is 1:1:0.5.

[0035] In certain embodiments, the step of dece...

Embodiment 1

[0064] Example 1 Decellularization of omentum derived from miniature pigs

[0065] Fresh minipig peritoneal omentum tissues (purchased from slaughterhouses) were taken and washed 3 times in PBS. The cleaned tissues were soaked in 2L of degreasing solution for 24 hours by shaking, and the shaking frequency was 200r / min. Subsequently, the fat-removed omentum tissue was washed three times in PBS, and soaked in 1L of decellularized solution I for 8 hours with shaking at a frequency of 150 r / min. Subsequently, the omentum tissue treated with decellularization solution I was washed 3 times in PBS, and soaked in 1L decellularization solution II for 8 hours with shaking at a frequency of 150r / min. Subsequently, the omentum tissue treated with decellularization solution II was washed 3 times in PBS, and soaked in 1L decellularization solution III for 8 hours with shaking at a frequency of 150r / min. Finally, the omentum tissue treated with the decellularization solution III was freeze...

Embodiment 2

[0066] Example 2 Immunohistochemical staining and identification of decellularized biological omentum

[0067] The decellularized biological omentum prepared in Example 1 was fixed with 4% paraformaldehyde solution, dehydrated with concentration gradient alcohol, embedded in paraffin, and sliced ​​with a thickness of 4 μm. The slices were dried on a pathological tissue drying apparatus for 2 hours. Subsequently, the tissue sections were hydrated with graded alcohol (from high to low concentration), and the sections were rinsed with distilled water. Add 3% hydrogen peroxide for 10 minutes to remove endogenous peroxidase; rinse three times with PBS, 5 minutes each time; use antigen retrieval solution for microwave repair, cool to room temperature after repair, rinse three times with PBS, 5 minutes each time; 1% Bovine serum albumin was blocked at 37°C for 30 minutes; anti-type I collagen (1:200), anti-IV collagen (1:200), anti-fibronectin antibody (1:200) and anti-elastin prima...

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Abstract

The invention discloses a preparation method of decellularized biological omentum and the decellularized biological omentum prepared therefrom. The preparation method of the decellularized biological omentum comprises the step of soaking the omentum with a buffer solution containing sodium dodecyl sulfate and DNase for decellularization. Also disclosed is the decellularized biological omentum prepared by the method. The method of the present invention combines gradient elution with physical shaking, has mild conditions, improves the decellularization efficiency of the omentum, and at the same time retains the composition of the acellular matrix of the omentum to the greatest extent; the method is also simple to operate and can be implemented strong advantage.

Description

technical field [0001] The invention relates to the technical field of tissue engineering and biomedical materials, in particular to a preparation method of decellularized biological omentum and the decellularized biological omentum prepared therefrom. Background technique [0002] The extracellular matrix (ECM) is a complex complex of structural and functional proteins, including a variety of fibrous proteins, such as collagen, fibronectin, laminin, and proteoglycan, and its components vary among different species. relatively conservative. The extracellular matrix acts as a scaffold for cell growth and plays an important role in cell adhesion, proliferation and differentiation. In the field of tissue engineering research, in order to better simulate the three-dimensional microenvironment of natural extracellular matrix, in recent years, researchers have developed a series of technologies and methods to obtain acellular matrix bioscaffold materials through decellularization...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/36A61L27/50
Inventor 饶义伟蓝德宾
Owner 北京奥泰康医药技术开发有限公司
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