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A Method for Efficiently Preparing Fat-derived Biomaterials Using Ultrasound

A biomaterial and ultrasonic technology, applied in the field of regenerative medicine, can solve the problems of cytokine loss, cumbersome operation, and long time consumption, and achieve the effect of improved cell activity, low preparation cost, and simple and easy method

Active Publication Date: 2022-02-18
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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Examples

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 fat-derived biological materials by using ultrasonic waves, which comprises the steps of centrifugation, ultrasonic crushing (including contact and non-contact ultrasonic waves), centrifugal collection and the like. The method of the present invention uses ultrasonic waves to break most of the mature fat cells in the fat tissue, and at the same time applies flocculent centrifugal sedimentation technology to gather the oil droplets produced by the broken mature fat cells, so as to achieve the purpose of enriching SVFs; and then use high-speed centrifugation to remove Cellular components, to obtain ECM. After ultrasonic treatment, the cell activity is greatly improved and the apoptosis rate is greatly reduced, which greatly improves the effectiveness of transplantation; at the same time, ECM biomacromolecular materials can be obtained, which avoids the low ECM content and Cytokine loss. The biological material prepared by the method of the invention can be used for autologous transplantation biotherapy, reduces the complication 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 stromal vascular fragment cell materials and extracellular matrix materials by utilizing 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 the treatment of various clinical diseases (Zuk, 2010). The study 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 are no domestic ESC products that meet the requirements of clinical research. In clinical treatment, the commonly used cell types are adult-derived cells, such as mesenc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61L27/36A61K35/35A61L27/60A61P17/02
Inventor 聂云飞
Owner 聂云飞
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