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Method for highly efficient separation of adipose mesenchymal stem cells from adipose extracts

A technology for adipose stem cells and preadipocytes, which is applied in the field of efficient separation of adipose-derived mesenchymal stem cells, can solve the problems of low acquisition efficiency of adipose-derived mesenchymal stem cells, poor state of adipose-derived mesenchymal stem cells, complicated and cumbersome separation process, etc. The method is simple and easy to implement, with strong repeatability and high acquisition rate.

Inactive Publication Date: 2019-04-02
PLASTIC SURGERY HOSPITAL CHINESE ACAD OF MEDICAL SCI
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Problems solved by technology

The separation process is complicated and costly, and there is no uniform operation standard, resulting in low efficiency and poor repeatability of adipose-derived mesenchymal stem cells, and the obtained adipose-derived mesenchymal stem cells are poor in state

Method used

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  • Method for highly efficient separation of adipose mesenchymal stem cells from adipose extracts
  • Method for highly efficient separation of adipose mesenchymal stem cells from adipose extracts
  • Method for highly efficient separation of adipose mesenchymal stem cells from adipose extracts

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Embodiment

[0041] A method for efficiently isolating adipose-derived mesenchymal stem cells from lipoaspirate, comprising the following steps:

[0042] Step 1: Obtaining lipoaspirate. Qualified hospitals obtain lipoaspirate through liposuction under aseptic conditions. The lipoaspirate is packaged in a sterile airtight syringe, stored at 4°C, and stored The time does not exceed 24h;

[0043]Step 2: Obtaining adipose tissue, washing the lipoaspirate obtained in step 1 with PBS, centrifuging the washed aspirate at a centrifugation rate of 1200rmp, the centrifugation time is 5min, and removing after centrifugation For the upper oil layer and the lower non-fat layer, absorb the fat tissue in the middle layer for later use, and record the volume of the obtained fat tissue;

[0044] Step 3: Acquire preadipocytes, add 0.2% type I collagenase solution to 10ml of the adipose tissue obtained in step 2, the volume of 0.1% type I collagenase solution is 10ml the same as that of adipose tissue, and ...

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Abstract

The invention relates to a method for highly efficient separation of adipose mesenchymal stem cells from adipose extracts, wherein the method comprises the following steps: step 1, obtaining of preadipocytes: adding a collagenase solution to an adipose tissue, shaking to obtain a mixed solution, adding a culture medium to the mixed solution, to obtain a secondary mixed solution, centrifuging the secondary mixed solution, and collecting bottom cells to obtain the preadipocytes; and step 2, obtaining of the adipose mesenchymal stem cells: resuspensing the preadipocytes obtained in the step 1 with a culture medium to obtain a cell resuspension, filtering the cell resuspension to remove impurities, to obtain a solution containing the adipose mesenchymal stem cells, and measuring the number ofcells in the solution containing the adipose mesenchymal stem cells per unit volume. The method for separation of the adipose mesenchymal stem cells from the adipose tissue is provided, wherein the method is simple in material obtaining and has quantitative steps and high repeatability.

Description

technical field [0001] The invention relates to the technical field of biological cell separation, in particular to a method for efficiently separating adipose-derived mesenchymal stem cells from lipoaspirate. Background technique [0002] Liposuction after liposuction is medical waste. After the operation, it needs to be treated as medical waste. However, a large number of fat stem cells are discarded together. A large number of adipose-derived stem cells (ADSCs) are stored in lipoaspirate. Like bone marrow mesenchymal stem cells and other human tissue-derived stem cells, ADSCs play an important role in tissue regeneration and tissue repair. The role of ADSC can be either a seed cell, which differentiates from a stem cell into a mature cell required for tissue regeneration and repair (such as adipocytes), or an inducer, which inhibits the surrounding inflammatory response and promotes tissue angiogenesis through paracrine function. Utilizing the above functions of ADSCs ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N5/077
CPCC12N5/0653C12N5/0667C12N2509/00
Inventor 李梓菲栾杰王黔穆大力刘春军辛敏强付苏
Owner PLASTIC SURGERY HOSPITAL CHINESE ACAD OF MEDICAL SCI
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