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Method for separating mesenchymal stem cells from dispose tissues

An adipose stem cell and adipose tissue technology, applied in the direction of bone/connective tissue cells, animal cells, vertebrate cells, etc., can solve the problems of strict specifications, lack of details of adipose stem cells, etc. repeatable effect

Active Publication Date: 2011-03-09
UNION STEMCELL & GENE ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there is no generally accepted strict standard for the specific method and some details of the separation of adipose stem cells.

Method used

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  • Method for separating mesenchymal stem cells from dispose tissues
  • Method for separating mesenchymal stem cells from dispose tissues

Examples

Experimental program
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Effect test

Embodiment 1

[0040] A total of 180ml of expanded liposuction was obtained from the hospital, and transported to the laboratory within 2 hours at 4°C after liposuction using a biological transport bag, and processed in a sterile state:

[0041] 1. Repeatedly wash with D-Hanks solution incubated at 37°C until the washing solution is colorless to ensure that the red blood cells are clean.

[0042] 2. Add collagenase type I prepared with D-Hanks solution to a final concentration of 0.1g / 100ml, and digest with shaking at 100 rpm at 37°C for 1 hour

[0043] 3. Centrifuge the digested product at 4°C and 450 g for 5 min; resuspend in 10 ml of adipose stem cell culture medium, filter with a 100 μm pore size filter to remove impurities, and finally obtain adipose stem cells.

[0044] 4. The adipose stem cell suspension and the red blood cell lysate were mixed at 1:1 and incubated for 2 minutes, centrifuged for 5 minutes at 4°C and a centrifugal force of 450 g, and the adipose stem cells were resuspe...

Embodiment 2

[0049] Adipose tissue is 180g of adipose tissue waste obtained during surgery. Using sterile reagent bottles, it is transported to the laboratory under aseptic conditions at 4°C and within 2 hours, and processed under aseptic conditions:

[0050] 1. First cut it into pieces as much as possible to about 1mm 3Large and small tissue blocks were repeatedly washed with D-Hanks solution incubated at 37°C until the washing solution was colorless to ensure that the red blood cells were cleaned.

[0051] 2. Add collagenase type I prepared with D-Hanks solution to a final concentration of 0.2g / 100ml, and digest with shaking at 100 rpm at 37°C for 1 hour

[0052] 3. Centrifuge the digested product at 4°C and 450 g for 5 min; resuspend in 10 ml of adipose stem cell medium, filter with a 100 μm pore size filter to remove impurities, and finally obtain adipose stem cells.

[0053] 4. The adipose stem cell suspension and the red blood cell lysate were mixed at 1:1 and incubated for 2 minute...

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Abstract

The invention relates to a method for separating mesenchymal stem cells from dispose tissues. The method comprises the following steps: putting dispose tissues to a biological bag for transportation or a sterilized reagent bottle; in aseptic conditions, removing red blood cells preliminarily; digesting dispose extract to obtain adipose-derived stem cells; removing red blood cells once again, and counting the adipose-derived stem cells; and carrying out activity detection to the adipose-derived stem cells, and cultivating the adipose-derived stem cells to obtain the mesenchymal stem cells. In the method, a large quantity of adipose-derived stem cells which are in good conditions and have good multi-difference capability can be obtained with a small quantity of dispose tissue. The method has the advantages of easiness and good reproducibility.

Description

technical field [0001] The present invention relates to a method for isolating mesenchymal stem cells, especially a method for isolating mesenchymal stem cells from adipose tissue. Background technique [0002] Since the discovery of adipose-derived stem cells in 2001, they have great advantages and broad prospects in clinical applications due to their large number and relatively simple access methods. The research of adipose stem cells is a hot spot in stem cell research and stem cell therapy, and related clinical treatments and clinical trials are underway. The researchers not only proved that it has the ability to differentiate into various mesodermal cells (including adipocytes, osteoblasts, chondrocytes and skeletal muscle cells), but also can be induced to become islet cells, nerve cells, cells of other germ layers such as hepatocytes. With the deepening of research, its clinical application value has attracted more and more attention. [0003] As researchers contin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775
Inventor 陈晓波韩洪起黄家学
Owner UNION STEMCELL & GENE ENG
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