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Method for efficiently preparing fat source biomaterial by using ultrasonic wave

A biomaterial and ultrasonic technology, applied in the field of regenerative medicine, can solve the problems of cytokine loss, cumbersome operation, and high cost, and achieve the effects of increased cell activity, low preparation cost, and improved survival rate

Active Publication Date: 2019-06-14
聂云飞
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] 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 efficiently preparing fat source biomaterial by using ultrasonic wave
  • Method for efficiently preparing fat source biomaterial by using ultrasonic wave

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0049] A method for efficiently preparing fat-derived biological materials by using ultrasonic waves, the specific steps are as follows:

[0050] (1) Centrifuge the adipose tissue at 5000rpm for 3 minutes. After centrifugation, if it is divided into three layers, namely the top layer of fat, the middle layer of mixture and the bottom layer of swelling fluid, discard the bottom layer of swelling fluid, filter to remove the top layer of fat and take the filtrate; if If there is no delamination or the delamination is not obvious, repeat this step;

[0051] (2) performing non-contact ultrasonic crushing on the filtrate obtained in step (1), the ultrasonic crushing conditions being 1000W, 10min, 36°C;

[0052] (3) centrifuging the fat mixture after step (2) non-contact ultrasonic crushing at 5000rpm for 5min;

[0053] (4) After centrifugation in step (3), if the fat mixture is divided into three layers, which are the top layer of fat, the middle layer of mixture and the bottom swe...

Embodiment 2

[0058] A method for efficiently preparing fat-derived biological materials by using ultrasonic waves, the specific steps are as follows:

[0059] (1) Centrifuge the adipose tissue at 5500rpm for 3 minutes. After centrifugation, if it is divided into three layers, namely the top layer of fat, the middle layer of mixture and the bottom layer of swelling fluid, discard the bottom layer of swelling fluid, filter to remove the top layer of fat and take the filtrate; if If there is no delamination or the delamination is not obvious, repeat this step;

[0060] (2) The filtrate obtained in step (1) is subjected to non-contact ultrasonic crushing, and the ultrasonic crushing conditions are 1100W, 9min, 36°C;

[0061] (3) centrifuging the fat mixture after step (2) non-contact ultrasonic crushing at 5000rpm for 6min;

[0062] (4) After centrifugation in step (3), if the fat mixture is divided into three layers, which are the top layer of fat, the middle layer of mixture and the bottom ...

Embodiment 3

[0067] A method for efficiently preparing fat-derived biological materials by using ultrasonic waves, the specific steps are as follows:

[0068] (1) Centrifuge the adipose tissue at 6000rpm for 3 minutes. After centrifugation, if it is divided into three layers, namely the top layer of fat, the middle layer of mixture and the bottom layer of swelling fluid, discard the bottom layer of swelling fluid, filter to remove the top layer of fat and take the filtrate; If there is no delamination or the delamination is not obvious, repeat this step;

[0069] (2) The filtrate obtained in step (1) is subjected to non-contact ultrasonic crushing, and the ultrasonic crushing conditions are 1150W, 5min, and 36.5°C;

[0070] (3) centrifuging the fat mixture after step (2) non-contact ultrasonic crushing at 5000rpm for 4min;

[0071] (4) After centrifugation in step (3), if the fat mixture is divided into three layers, which are the top layer of fat, the middle layer of mixture and the bott...

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Abstract

The invention discloses a method for efficiently preparing a fat source biomaterial by using ultrasonic wave. The method comprises the steps of centrifugation, ultrasonic wave breaking (including contact type and non-contact type ultrasonic wave), centrifugal collection and the like. According to the method, the ultrasonic wave is utilized to break most mature fat cells in fat tissues, and the flocculation centrifugal sedimentation technology is applied to collect oil droplets generated by breaking the mature fat cells to achieve the purpose of enriching SVFs; high-speed centrifuging is utilized to remove cellular components to obtain ECM. After ultrasonic treatment, the cell activity is greatly improved, and the apoptosis rate is greatly reduced, so that the effectiveness of the transplantation is greatly improved. At the same time, an ECM biomacromolecule material can be obtained, and the low ECM content and the cytokine loss caused by the cumbersome operation are avoided. The biomaterial obtained by the method can be used for the autologous transplantation biological therapy, reduces the complications of fat transplantation and improves the survival rate of transplanted fat.

Description

technical field [0001] The invention relates to the technical field of regenerative medicine, in particular to a method for efficiently preparing fat-derived biological materials by using ultrasonic waves. More specifically, it relates to a method for efficiently preparing cell materials and extracellular matrix materials from stromal vessel fragments by using contact and non-contact ultrasonic waves. Background technique [0002] Stem cells can survive for a long time, have continuous self-reproduction ability and multi-directional potential, and provide new ideas for clinical treatment of various diseases (Zuk, 2010). Studies have found that there are two main sources of stem cells, namely, embryonic stem cells (mesenchymal stem cells, ESCs) and adult stem cells (Adult stem cells, ASCs). At present, there is no domestic ESC product that meets the clinical research requirements. In clinical treatment, the commonly used cell types are adult-derived cells, such as mesenchyma...

Claims

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Application Information

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IPC IPC(8): A61L27/36A61K35/35A61L27/60A61P17/02
Inventor 聂云飞张玲华
Owner 聂云飞
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