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Preparation method for efficiently enriching SVF (stromal vascular fraction) cell material through built-in ultrasonic waves

A stromal blood vessel and ultrasonic technology, applied in the field of regenerative medicine, can solve the problems of high loss of dry glue cells, complicated separation process, cumbersome crushing process, etc., and achieve the effects of decreased cell apoptosis rate, low preparation cost, and simple and easy method

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] However, there are still many limiting factors in the stem cell therapy of adipose-derived stem cells and stromal vascular fragment cells. For example, the effectiveness of post-transplantation is controversial, and the isolation process is complicated. The existing isolation methods mainly include enzymatic hydrolysis, bolus crushing, etc.
However, the enzymatic hydrolysis method adds exogenous digestive enzymes in the enrichment process, and it is expensive and needs to be carried out under specific experimental conditions; The glue may contain small amounts of glass shards, potentially causing inflammation

Method used

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  • Preparation method for efficiently enriching SVF (stromal vascular fraction) cell material through built-in ultrasonic waves

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Effect test

Embodiment 1

[0034] A preparation method for efficiently enriching stromal vessel fragment cell material by using built-in ultrasonic waves, the specific steps are as follows:

[0035] (1) Centrifuge the obtained adipose tissue at 5000rpm for 3min, and discard the bottom blood after centrifugation (when pushing out the swelling fluid in the lower layer, the swelling fluid should be pushed as clean as possible without losing fat, otherwise the final product will contain blood color), and then passed through a 60-mesh sieve (after filtration, the adipose tissue remaining in the filter should be sucked under negative pressure to avoid excessive fat loss), and the filtrate was transferred to a 15mL centrifuge tube (when transferring the upper and middle layer mixture, try not to Transfer the upper layer of fat into a centrifuge tube);

[0036] (2) Adjust the frequency of the ultrasonic cell disruptor to 75W, insert the probe of the ultrasonic cell disruptor into the centrifuge tube filled with...

Embodiment 2

[0040] A preparation method for efficiently enriching stromal vessel fragment cell material by using built-in ultrasonic waves, the specific steps are as follows:

[0041] (1) Centrifuge the obtained adipose tissue at 5000rpm for 3min, and discard the bottom blood after centrifugation (when pushing out the swelling fluid in the lower layer, the swelling fluid should be pushed as clean as possible without losing fat, otherwise the final product will contain blood color), and then passed through a 60-mesh sieve (after filtration, the adipose tissue remaining in the filter should be sucked under negative pressure to avoid excessive fat loss), and the filtrate was transferred to a 15mL centrifuge tube (when transferring the upper and middle layer mixture, try not to Transfer the upper layer of fat into a centrifuge tube);

[0042] (2) Adjust the frequency of the ultrasonic cell disruptor to 70W, insert the probe of the ultrasonic cell disruptor into the centrifuge tube filled with f...

Embodiment 3

[0046] A preparation method for efficiently enriching stromal vessel fragment cell material by using built-in ultrasonic waves, the specific steps are as follows:

[0047] (1) Centrifuge the obtained adipose tissue at 5000rpm for 3min, and discard the bottom blood after centrifugation (when pushing out the swelling fluid in the lower layer, the swelling fluid should be pushed as clean as possible without losing fat, otherwise the final product will contain blood color), and then passed through a 60-mesh sieve (after filtration, the adipose tissue remaining in the filter should be sucked under negative pressure to avoid excessive fat loss), and the filtrate was transferred to a 15mL centrifuge tube (when transferring the upper and middle layer mixture, try not to Transfer the upper layer of fat into a centrifuge tube);

[0048] (2) Adjust the frequency of the ultrasonic cell disruptor to 65W, insert the probe of the ultrasonic cell disruptor into the centrifuge tube filled with...

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Abstract

The invention belongs to the technical field of regenerative medicine and discloses a preparation method for efficiently enriching a SVF (stromal vascular fraction) cell material 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, and meanwhile, oil drops produced by crushed mature adipose cells are collected with a floccus centrifugal precipitation technology, so that the effect of enriching SVFs is realized. After ultrasonic treatment, the cell viability is substantially improved and the cell apoptosis rate is substantially decreased, so that the transplanting effectiveness is greatly enhanced. The cell 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 invention relates to the technical field of regenerative medicine, in particular to a preparation method for efficiently enriching stromal blood vessel fragment cell materials by using built-in ultrasonic waves. Background technique [0002] Adipose tissue is mainly composed of adipocytes, adipose-derived stromal / stemcells (ASCs), vascular endothelial cells, pericytes, fibroblasts, macrophages, and extracellular matrix. ASCs are a group of cells with multi-directional differentiation potential. Vascular stromal fraction (SVFs) in adipose tissue is obtained after adipose tissue is digested with collagenase, filtered, and centrifuged to remove mature adipocytes. It contains abundant and highly plastic ASCs , including stromal cells, vascular endothelial cells, vascular smooth muscle cells, pericytes, immune cells and a large number of cells from blood circulation, such as white blood cells, red blood cells, etc., which are the most important componen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61L27/36A61L27/60A61K8/98A61Q19/08
CPCA61L27/3604A61K8/981A61L27/3691A61L27/60A61L2430/34A61Q19/08
Inventor 聂云飞卢沛伶张玲华
Owner 聂云飞
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