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Method for preparing adipose-derived ECM (extracellular matrix) through built-in ultrasonic waves

A technology of adipose-derived cells and extracellular matrix, applied in the field of preparation of adipose-derived extracellular matrix by built-in ultrasound, can solve the problems of cytokine loss, cumbersome operation, high cost, etc., achieve low preparation cost, increase survival rate, and broad application prospects Effect

Inactive Publication Date: 2018-11-06
聂云飞
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] The current method of separating ECM is mainly the method of repeated freezing and thawing + multiple enzymatic hydrolysis + polar solvent extraction proposed by Flynn. , and it needs to be carried out under specific experimental conditions, so it is not of great clinical value

Method used

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  • Method for preparing adipose-derived ECM (extracellular matrix) through built-in ultrasonic waves

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] A method for preparing fat-derived extracellular matrix using built-in ultrasound, the specific steps are as follows:

[0034] (1) Centrifuge the obtained adipose tissue at 5000rpm for 2min, and discard the bottom blood after centrifugation (when pushing out the lower layer of swelling fluid, push the swelling fluid as clean as possible without losing fat, otherwise the final product will have Blood color), then pass through a 70-mesh screen (after filtering, the adipose tissue remaining in the filter should be sucked under negative pressure to avoid losing too much fat), and the filtrate is transferred to a sterile centrifuge tube (when transferring the middle and upper layer mixture, as much as possible Do not transfer the upper layer of grease into the centrifuge tube);

[0035] (2) Put the fat tissue into the ultrasonic cell disruptor, adjust the power to 45W and time for 25s for ultrasonic disruption; then continue ultrasonic disruption, the ultrasonic disruption condit...

Embodiment 2

[0039] A method for preparing fat-derived extracellular matrix using built-in ultrasound, the specific steps are as follows:

[0040] (1) Centrifuge the obtained adipose tissue at 4000 rpm for 5 minutes, and discard the bottom blood after centrifugation (when pushing out the lower swelling fluid, push the swelling fluid as clean as possible without losing fat, otherwise the final product will have Blood color), then pass through a 60-mesh screen (after filtering, the adipose tissue remaining in the filter should be sucked under negative pressure to avoid losing too much fat), and the filtrate is transferred to a sterile centrifuge tube (when transferring the middle and upper layer mixture, as much as possible Do not transfer the upper layer of grease into the centrifuge tube);

[0041] (2) Put the fat tissue into the ultrasonic cell disruptor, adjust the power to 50W, and time for 15s for ultrasonic disruption; then continue ultrasonic disruption, the ultrasonic disruption conditio...

Embodiment 3

[0045] A method for preparing fat-derived extracellular matrix using built-in ultrasound, the specific steps are as follows:

[0046] (1) Centrifuge the obtained adipose tissue at 4000 rpm for 5 minutes, and discard the bottom blood after centrifugation (when pushing out the lower swelling fluid, push the swelling fluid as clean as possible without losing fat, otherwise the final product will have Blood color), then pass through an 80-mesh screen (after filtering, the adipose tissue remaining in the filter should be sucked under negative pressure to avoid losing too much fat), and the filtrate is transferred to a sterile centrifuge tube (when transferring the middle and upper mixture, as much as possible Do not transfer the upper layer of grease into the centrifuge tube);

[0047] (2) Put the fat tissue into the ultrasonic cell disruptor, adjust the power to 65W, and time for 10s to perform ultrasonic disruption; then continue ultrasonic disruption, the ultrasonic disruption condit...

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Abstract

The invention belongs to the technical field of regenerative medicine and discloses a method for preparing an adipose-derived ECM (extracellular matrix) through built-in ultrasonic waves, which comprises steps of centrifugation, ultrasonic crushing, centrifugal collection and the like. According to the preparation method, most mature adipose cells in adipose tissue are crushed through ultrasonic waves, manure adipose cells and SVF cells are removed, the ECM is prepared, and too low ECM content and cell factor loss caused by complex operation are avoided effectively. According to the method, the ECM rich in cell factors is prepared without addition of exogenous chemical substances, the prepared ECM contains no exogenous substances, and adipose autotransplantation and effectiveness of the ECM as a biological filling material can be improved. The adipose-derived ECM material prepared with the method can be applied to autotransplantation stem cell therapy, reduces complications caused by adipose transplantation, increases the survival rate of transplanted adipose and has broad application prospects.

Description

Technical field [0001] The present invention relates to the technical field of regenerative medicine, in particular to a method for preparing fat-derived extracellular matrix using built-in ultrasound. Background technique [0002] Fat transplantation is usually used by plastic surgery to solve contour abnormalities and soft tissue defects. However, the retention rate of fat cells and the effect of volume filling are unpredictable, so it is necessary to seek a new biological material. Preliminary studies on the use of extracellular matrix in human adipose tissue show that it has a unique role in fat transplantation. Under normal circumstances, a large amount of adipose tissue is discarded as medical waste during abdominoplasty, liposuction, body shaping and breast reduction surgery. So far, research focuses on the optimization of fat graft acquisition and processing, or its cellular components such as stromal vascular fragments (SVF). However, the extracellular matrix (ECM) of ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61K35/35A61L27/60A61P17/02
CPCA61L27/3633A61K35/35A61L27/3691A61L27/60A61L2430/34A61P17/02
Inventor 聂云飞张玲华
Owner 聂云飞
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