Separating, culturing and preparation preparing method for adipose tissue-derived stromal cells

A technology for the isolation and cultivation of mesenchymal stem cells, which is applied in the field of separation and cultivation of adipose-derived mesenchymal stem cells and the preparation of preparations, and can solve the problem of the introduction of mycoplasma, virus and other pathogenic microorganisms from porcine, exogenous pollution, and the introduction of pathogenic microorganisms such as mycoplasma and viruses from bovine Pollution and other issues, to achieve the effect of combined application is superior, high safety, improve the effect of skin quality

Pending Publication Date: 2019-04-23
王振坤
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The in vitro culture of mesenchymal stem cells has the risk of exogenous contamination. For example, the use of bovine serum in the culture system may introduce the risk of contamination by pathogenic microorganisms such as mycoplasma bovis and viruses, and the use of porcine trypsin in the culture system may introduce mycoplasma suis. The risk of contamination by pathogenic microorganisms such as viruses and viruses

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0037] 1) Collect 60% of peripheral blood in EDTA anticoagulant tubes, and transfer them to two 50ml centrifuge tubes in a biological safety cabinet. Centrifuge at 700g for 10 minutes, absorb the upper layer of yellow plasma into a clean 50ml centrifuge tube, add normal saline to the remaining blood to the original volume.

[0038] 2) Add 15ml of Percoll with a density of 1.057 to two 50ml centrifuge tubes, then slowly add 30ml of the above-mentioned peripheral blood diluted with normal saline, and centrifuge at 500g for 20 minutes.

[0039] 3) Aspirate the platelets in the middle and upper layers, put them into a clean 50ml centrifuge tube, and centrifuge at 400g for 8 minutes.

[0040] 4) Transfer the supernatant to a clean 50ml centrifuge tube and centrifuge at 1600g for 5 minutes.

[0041] 5) Discard the supernatant, and take the plasma in step 1 to resuspend the platelet pellet.

[0042] 6) The centrifuge tube containing the platelet-rich plasma was placed in liquid nit...

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PUM

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Abstract

The invention relates to the field of adipose-tissue-derived-stromal-cell medical cosmetology, and discloses a separating, culturing and preparation preparing method for adipose tissue-derived stromalcells. According to the method, autologous adipose tissue is treated with the physical method, collagenase does not need to be introduced, long-time tedious treatment does not need, and extracellularmatrixes are reserved and fully used; autologous adipose tissue-derived stromal cells are further amplified through autologous platelet-factor-rich plasma, and sufficient autologous adipose tissue-derived stromal cells and stem cell factors are obtained; the adipose tissue-derived stromal cells and the extracellular matrixes have the good effect on rhytidectomy, soft tissue repairing and facial rejuvenation, and the stem cell factors can be widely applied to skin repairing.

Description

technical field [0001] The invention relates to a method for separating, culturing and preparing adipose-derived mesenchymal stem cells. The invention belongs to the medical cosmetology field of adipose mesenchymal stem cells. Background technique [0002] Stem cells are a type of pluripotent cells with self-renewal and self-replication capabilities. Mesenchymal stem cells are an important member of the stem cell family, derived from the mesoderm in the early stage of development, and can differentiate into fat, bone, cartilage, muscle, tendon, ligament, nerve, liver, cardiac muscle, Various tissue cells such as endothelium are ideal seed cells for regenerative medicine. Mesenchymal stem cells exist in bone marrow, umbilical cord blood and umbilical cord tissue, placental tissue, adipose tissue and other tissues. [0003] Adipose-derived mesenchymal stem cells are a kind of stem cells isolated from adipose tissue in recent years with multipotential differentiation potenti...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C07K14/475C07K1/14
CPCC12N5/0667C07K14/475C12N2500/84C12N2509/10
Inventor 王振坤罗赛朱轶刘小龙
Owner 王振坤
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