Additive for preserving adipose-derived mesenchymal stem cells and application of additive

A technology of high-quality stem cells and additives, applied in the field of stem cells, can solve the problems of increased cost, protein denaturation, and high equipment requirements, and achieve the effects of safe ingredients, good proliferation activity, and stable storage environment

Active Publication Date: 2022-06-03
GUANGDONG CELL BIOTECHNOLOGY CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, dimethyl sulfoxide has a certain toxic effect, which can interact with protein hydrophobic groups, leading to protein denaturation, and has vascular toxicity and liver and kidney toxicity.
In addition, storing cells in liquid nitrogen requires high equipment requirements. As the storage time prolongs, liquid nitrogen needs to be replenished frequently, which increases the cost of storage.

Method used

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  • Additive for preserving adipose-derived mesenchymal stem cells and application of additive
  • Additive for preserving adipose-derived mesenchymal stem cells and application of additive

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] An additive for the preservation of adipose mesenchymal stem cells, the mass concentration of the additive in a low-sugar DMEM medium is: mannitol 2 mg / mL, silk fibroin 10 μg / mL, docusate sodium 8 μg / mL, glutathione 3 μg / mL mL, glycerol 0.8mg / mL, quercetin 12ng / mL.

[0025] The application of the above-mentioned additive for preservation of adipose-derived mesenchymal stem cells includes the following steps:

[0026] (1) adding additives to the culture medium, mixing to obtain a preservation solution, for subsequent use;

[0027] (2) Add the adipose-derived mesenchymal stem cells to be preserved in the preservation solution of step (1) to resuspend, and distribute them in 2 mL cryopreservation tubes. The density of adipose-derived mesenchymal stem cells in the preservation solution is 1×10 7 pcs / mL;

[0028] (3) Program cooling of the preservation solution in step (2) to -80°C, and the program cooling process is: cooling to -20°C at 1°C / min, and then cooling to -80°C ...

Embodiment 2

[0030] An additive for preservation of adipose mesenchymal stem cells, the mass concentration of the additive in a low-sugar DMEM medium is: mannitol 1 mg / mL, silk fibroin 7 μg / mL, docusate sodium 5 μg / mL, glutathione 1 μg / mL mL, glycerol 0.5mg / mL, quercetin 10ng / mL.

[0031] The application of the above-mentioned additive for preservation of adipose-derived mesenchymal stem cells includes the following steps:

[0032] (1) adding additives to the culture medium, mixing to obtain a preservation solution, for subsequent use;

[0033] (2) Resuspend the adipose-derived mesenchymal stem cells to be preserved with the preservation solution of step (1), and distribute them in 2 mL cryopreservation tubes. The density of adipose-derived mesenchymal stem cells in the preservation solution is 2×10 7 pcs / mL;

[0034] (3) Program cooling of the preservation solution in step (2) to -80°C, and the program cooling process is: cooling to -20°C at 2°C / min, and then cooling to -80°C at 18°C / mi...

Embodiment 3

[0036] An additive for preservation of adipose mesenchymal stem cells. mL, glycerol 1mg / mL, quercetin 15ng / mL.

[0037] The application of the above-mentioned additive for preservation of adipose-derived mesenchymal stem cells includes the following steps:

[0038] (1) adding additives to the culture medium, mixing to obtain a preservation solution, for subsequent use;

[0039] (2) Resuspend the adipose-derived mesenchymal stem cells to be preserved with the preservation solution of step (1), and distribute them in 2 mL cryopreservation tubes. The density of adipose-derived mesenchymal stem cells in the preservation solution is 3×10 7 pcs / mL;

[0040] (3) Program cooling of the preservation solution in step (2) to -80°C, and the program cooling process is as follows: cooling to -20°C at 3°C / min, and then cooling to -80°C at 20°C / min for preservation.

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Abstract

The invention discloses an additive for preserving adipose tissue-derived stem cells. The additive comprises the following components: mannitol, silk fibroin, docusate sodium, glutathione, glycerol and quercetin. All the components in the additive have a synergistic effect, the adipose-derived mesenchymal stem cells are preserved at the temperature of-80 DEG C, the components are safe, the preservation environment is stable, the survival rate of the resuscitated adipose-derived mesenchymal stem cells preserved for three months is 90% or above, and liquid nitrogen is not needed in the preservation process. The adipose-derived stem cells preserved by the additive are high in survival rate and good in proliferation activity after being recovered. The invention further provides specific application of the additive, the additive is added into a culture medium to be used for preserving the adipose-derived mesenchymal stem cells, operation is convenient, preservation conditions are easy to achieve, and the preservation cost of the adipose-derived mesenchymal stem cells is reduced.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to an additive for preservation of adipose mesenchymal stem cells and its application. Background technique [0002] Mesenchymal stem cells can be isolated from various tissues, such as bone marrow, adipose tissue, skin, skeletal muscle, and umbilical cord blood. Among them, stem cells derived from adipose tissue are called adipose-derived mesenchymal stem cells. The yield of adipose-derived mesenchymal stem cells is higher than that of bone marrow-derived mesenchymal stem cells. And it is rich in growth factors and cytokines, and has a very broad application prospect in the fields of cell repair, developmental biology, and pharmacology. Adipose-derived mesenchymal stem cells have been widely used in clinical cell therapy and various cosmetic and skin care products. They are of great significance for the study of the self-renewal mechanism of stem cells, the regulation of stem cell signa...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/021
Inventor 尹利民常永华
Owner GUANGDONG CELL BIOTECHNOLOGY CO LTD
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