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Method of induced differentiation of rat bone marrow mesenchymal stem cells into adipocytes

A bone marrow mesenchymal and adipocyte technology, applied in the field of animal experiments, can solve the problems of long time and low induction differentiation rate, and achieve the effect of fast induction speed, simple configuration, and high induction rate

Inactive Publication Date: 2016-07-20
张君
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Problems solved by technology

[0009] The purpose of the present invention is to provide a method for inducing differentiation of rat bone marrow mesenchymal stem cells into adipocytes, aiming to solve the problem of low induction differentiation rate and long time in traditional culture methods

Method used

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  • Method of induced differentiation of rat bone marrow mesenchymal stem cells into adipocytes
  • Method of induced differentiation of rat bone marrow mesenchymal stem cells into adipocytes
  • Method of induced differentiation of rat bone marrow mesenchymal stem cells into adipocytes

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Embodiment 1

[0052] Two Wista male rats, 4 weeks old, were purchased from the Experimental Animal Center of Xinjiang Medical University. The handling of the animals during the experiment was in accordance with the regulations of the relevant animal ethics standards of "Ethical issues in animal experimentation" in 2009.

[0053] PBS phosphate buffer saline powder was purchased from Beijing Zhongshan Jinqiao Biotechnology Co., Ltd., fetal bovine serum, DMEM medium, 0.25% trypsin, penicillin-streptomycin double antibody were purchased from American Lifetechnologies company, dexamethasone, indomethacin , IBMX, and ascorbic acid were purchased from Sigma in the United States, insulin was purchased from Novo Nordisk Pharmaceutical Co., Ltd., oil red O staining kit was purchased from Nanjing Jiancheng Technology Co., Ltd., Anti-Mouse CD29, CD44 were purchased from eBioscience, and oleic acid was purchased from Shanghai .

[0054] (1) Traditional method to induce adipogenic differentiation

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Abstract

The invention discloses a method of induced differentiation of rat bone marrow mesenchymal stem cells into adipocytes, which includes: choosing and inoculating third-generation bone marrow mesenchymal stem cells into a cell culture flask according to a concentration of 1 multiplied by 106mL<-1>, and after the cells attaching to a wall grow until cell density reaches 80 percent, adding different concentrations of oleic acid as inducers; according to a volume proportion of 10 percent, adding fetal bovine serum into a basic culture medium DMEM (Dulbecco modified Eagle medium), and respectively adding 25mM of oleic acid, 50mM of oleic acid, 100mM of oleic acid and 200mM of oleic acid as induction culture media for forming adipocytes; replacing the solution once every three days, and with a cell culture flask without a induction culture solution as a control, performing oil red O staining assay after 3 weeks. According to the method, different concentrations of oleic acid can be used to directly induce adipocytes, a novel induction method is established, the configuration of the method is simpler, the induction speed is high, and the induction rate is high. In addition, the method can be used for inducing adipocytes of different sizes and researching the functional differences of the adipocytes of different sizes.

Description

technical field [0001] The invention belongs to the field of animal experiments, and in particular relates to a method for inducing differentiation of rat bone marrow mesenchymal stem cells into adipocytes. Background technique [0002] At present, domestic and foreign scholars adopt different adipogenic induction prescriptions, Zhuang Shubo et al. [1] The adipogenic inducer used in the relevant experiments was 1 μmol / L dexamethasone, 0.5 mmol / L 3-isobutylmethylxanthine, 1 mol / L human insulin, 1 mmol / L indomethacin, and the volume fraction was 10% fetal bovine Serum induction medium. Li Shengfu et al [2] Induce adipogenic differentiation of rabbit mesenchymal stem cells with adipogenic inducer DMEM induction medium (dexamethasone 10-8mol / L, insulin 10mg / L, penicillin 100U / mL, streptomycin 100mg / L, volume fraction 10% Fetal bovine serum, pH 7.2), showed that insulin and dexamethasone play an important role in the differentiation of adipocytes and are its initiation and pro...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/0775C12N5/077
CPCC12N5/0653C12N2501/999C12N2506/1346
Inventor 张君李伟王翠喆谢建新许彭哈晓丹顾雅娟
Owner 张君
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