Liver acellular stent construction method based on irreversible electroporation technology

A technology of decellularized scaffolds and construction methods, which is applied in the direction of cell dissociation methods, artificial cell constructs, animal cells, etc., can solve the problems of limited application range and long reaction time, and achieve wide application range, short time consumption, and damage small effect

Pending Publication Date: 2020-08-11
THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
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Problems solved by technology

However, in this study, the researchers mainly used the body's own immune response to achieve decellularization. The reaction takes a long time and can only be carried out in vivo, which limits its application range.

Method used

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  • Liver acellular stent construction method based on irreversible electroporation technology
  • Liver acellular stent construction method based on irreversible electroporation technology
  • Liver acellular stent construction method based on irreversible electroporation technology

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Embodiment Construction

[0032] see figure 1 , the method step of the specific embodiment of the present invention is as follows:

[0033] 1) Preparation of rat liver decellularized scaffolds.

[0034] 1.1) Animal preparation: the animals were weighed, anesthetized by gas inhalation with isoflurane and maintained, after the anesthesia took effect, SD rats were placed in a supine position, the abdominal skin was disinfected with 75% alcohol, and the liver was exposed by layered laparotomy;

[0035] 1.2) Irreversible electroporation of the liver: the left outer lobe of the rat liver was clamped with 1cm caliper electrodes, and treated with a high-voltage pulsed electric field (processing parameters: 2000V / cm, 90μs, 70 pulses, 1HZ);

[0036] 1.3) In vivo perfusion via portal vein: After the irreversible electroporation treatment, intubate through the portal vein of rats, perfuse with 100 U / mL heparinized PBS solution at a rate of 5 mL / min, and cut the subhepatic inferior vena cava at the same time, See...

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Abstract

The invention relates to a liver acellular stent construction method based on an irreversible electroporation technology. The method comprises the following steps of 1.1) weighing a rat, carrying outgas inhalation anesthesia by using isoflurane, maintaining the gas inhalation anesthesia, after the anesthesia is effective, disinfecting abdominal skin of the SD rat in a supine position by using 75%alcohol, and opening the abdomen layer by layer to expose the liver; 1.2) clamping a left lateral lobe area of the rat liver by using a caliper type electrode, and performing high-voltage pulsed electric field treatment; 1.3) after the irreversible electroporation treatment is finished, performing transhepatic portal vein catheterization, perfusing with heparinized PBS (Phosphate Buffer Solution), cutting off the subhepatic inferior vena cava until the liver is full and the color is gradually lightened, gradually dissociating tissues and ligaments around the liver, taking down the liver, andimmersing the liver into the heparinized PBS; 1.4) putting the taken liver into a clean container, and performing one-way perfusion with deionized water, and 1.5) after the deionized water perfusion is finished, performing one-way perfusion with PBS for 30 minutes to complete the preparation of the stent. By the method disclosed by the invention, the construction efficiency of the in-vitro acellular stent can be effectively improved under the condition of not applying harmful chemical preparations.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a method for constructing liver decellularized scaffolds based on irreversible electroporation technology. Background technique [0002] In recent years, the use of tissue engineering methods to prepare decellularized organ matrix scaffolds, retaining the organ's complete extracellular matrix components and natural vascular system, and constructing functional organs to solve the serious shortage of organs is a new trend in tissue engineering research. breakthroughs and hotspots. At present, biological decellularized scaffolds have been successfully applied to the repair and reconstruction research and clinical application of homogeneous or heterogeneous donor bladder, esophagus, trachea, skin, heart valve, heart, liver, lung and other tissues and organs. In 2008, Harald COtt's team first reported the rat heart decellularized scaffold, and constructed isolated cardiac tissue with cer...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/071A61L27/36
CPCC12N5/067A61L27/3604A61L27/3691C12N2509/00A61L2430/40A61L2430/28
Inventor 吕毅李青山任冯刚陈雪张雨驰畅朋康陈伯文王博吴荣谦马锋
Owner THE FIRST AFFILIATED HOSPITAL OF MEDICAL COLLEGE OF XIAN JIAOTONG UNIV
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