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Application and method for promoting adipose-derived mesenchymal stem cell proliferation and chondrogenic differentiation through b-D-Glucopyranoside

A technology of mesenchymal stem cells and eucommia glucoside, which is applied in the field of eucommia glucoside to promote the proliferation and chondrogenic differentiation of adipose-derived mesenchymal stem cells. It can solve the problems of not being widely used, expensive, and safety issues, and achieve the promotion of chondrogenic differentiation and low cost. , significantly improved effect

Active Publication Date: 2021-11-02
GUANGDONG SECOND TRADITIONAL CHINESE MEDICINE HOSPITAL (GUANGDONG PROVINCE ENG TECH RES INST OF TCM)
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In previous studies, some chemical substances and cytokines were often used to promote the proliferation and chondrogenic differentiation of adipose-derived mesenchymal stem cells, but they could not be widely used in clinical practice due to high price and safety issues.

Method used

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  • Application and method for promoting adipose-derived mesenchymal stem cell proliferation and chondrogenic differentiation through b-D-Glucopyranoside
  • Application and method for promoting adipose-derived mesenchymal stem cell proliferation and chondrogenic differentiation through b-D-Glucopyranoside
  • Application and method for promoting adipose-derived mesenchymal stem cell proliferation and chondrogenic differentiation through b-D-Glucopyranoside

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Embodiment Construction

[0026] The principles and features of the present invention will be described below in conjunction with examples and accompanying drawings, and the examples given are only used to explain the present invention, and are not intended to limit the scope of the present invention.

[0027] 1. Experimental equipment:

[0028] Cellometer Mini automatic cell counter (U.S. Nexcelom company);

[0029] Varioskan Flash full-wavelength multifunctional microplate reader (Thermo, USA);

[0030] 3K30 refrigerated high-speed centrifuge (Germany Sigma);

[0031] Class II Type B2 biological safety cabinet (Singapore Esco Company);

[0032] 3111CO 2 Cell incubator (U.S. Thermo company);

[0033] DMI8 inverted fluorescence microscope (Leica, Germany);

[0034] StepOnePlus real-time fluorescent quantitative PCR instrument (Thermo Company, USA);

[0035] 40 μm cell strainer (Biologix, USA).

[0036] 2. Experimental steps:

[0037] S1. Obtaining primary adipose-derived mesenchymal stem cells:...

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Abstract

The invention discloses application and a method for promoting adipose-derived mesenchymal stem cell proliferation and chondrogenic differentiation by b-D-Glucopyranoside. The method comprises the following steps: S1, acquiring primary adipose-derived mesenchymal stem cells; S2, culturing the primary adipose-derived mesenchymal stem cells in vitro; S3, promoting proliferation: digesting second-generation full adipose-derived mesenchymal stem cell with 0.25% trypsin, when the cells grow and the fusion degree reaches 80%, adding 10ng / mL eucommia ulmoides glycoside into the basic culture medium, and continuously culturing in a 5% CO2 incubator at 37 DEG C for 24 hours; S4, promoting chondrogenic differentiation: taking second-generation full adipose-derived mesenchymal stem cells, digesting the adipose-derived mesenchymal stem cell with 0.25% trypsin, adding 10 ng / mL of b-D-Glucopyranoside, 10 ng / mL of TGF-beta3, 100 nmol / L of dexamethasone, 1% of ITS-G, 50 [mu] g / mL of vitamin C and 40 [mu] g / mL of proline into a basic culture medium, changing the liquid once every 2 days, and continuously performingculturing for 14 days. According to the method, proliferation and chondrogenic differentiation of the adipose-derived stem cells are promoted, meanwhile, the cost can be reduced, and the safety is good.

Description

technical field [0001] The invention belongs to the technical field of stem cells, and in particular relates to the application and method of Eucommia glucoside for promoting the proliferation and chondrogenic differentiation of adipose mesenchymal stem cells. Background technique [0002] In recent years, with the in-depth research on adipose-derived mesenchymal stem cells, its prospects in the treatment of knee osteoarthritis have become more prominent. Promoting the proliferation of adipose-derived mesenchymal stem cells and improving their chondrogenic differentiation ability has always been a research hotspot and difficulty. In previous studies, some chemical substances and cytokines were often used to promote the proliferation and chondrogenic differentiation of adipose-derived mesenchymal stem cells, but they could not be widely used in clinical practice due to high price and safety issues. Contents of the invention [0003] Aiming at the above-mentioned problems e...

Claims

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Application Information

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IPC IPC(8): C12N5/077C12N5/0775
CPCC12N5/0667C12N5/0655C12N2500/76
Inventor 姚楠胡子旋刘文刚黄丹娥黄雪君陈玉兴
Owner GUANGDONG SECOND TRADITIONAL CHINESE MEDICINE HOSPITAL (GUANGDONG PROVINCE ENG TECH RES INST OF TCM)
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