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Method for preparing glioma cell senescence model by using D-galactose

A glioma, cell aging technology, applied in tumor/cancer cells, preparation of test samples, animal cells, etc., can solve the problems of low success rate, time-consuming and labor-intensive, reagent toxicity, etc., and achieve high success rate , good effect, easy to operate effect

Active Publication Date: 2020-02-21
HANGZHOU NORMAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0005] The purpose of the present invention is to overcome the defects that the methods used in the prior art for preparing tumor cell senescence models are usually time-consuming, labor-intensive, cost-intensive, and have a low success rate, and the reagents used at the same time are highly toxic and often cause a large number of cell death. , providing a simple, effective, low-cost method for preparing a senescence model of glioma cells using D-galactose without causing massive cell death

Method used

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  • Method for preparing glioma cell senescence model by using D-galactose
  • Method for preparing glioma cell senescence model by using D-galactose
  • Method for preparing glioma cell senescence model by using D-galactose

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Embodiment 1

[0040] In order to prepare the senescence of glioma cells, we chose rat glioma cells --- C6 cells. Place C6 cells at 37°C, 5% CO 2 cultured in an incubator. The medium is DMEM + 10% FBS + 1% PS. After the cells are congested, use 1×10 5 The density was inoculated, and when the cells grew to about 70%, they were treated with 40g / L D-galactose. Change the medium every other day to prevent D-galactose from becoming invalid. On days 2, 4, 6, and 8, the cells growing on the glass slides were harvested respectively. When the drug treatment time exceeds 3 days, subculture is carried out, a part of the cells are transferred to the slide, and another part of the cells are transferred to the culture dish without the slide. And so on until the 8th day sample is collected.

[0041] Since SA-β-Gal staining is the gold standard for judging cell senescence, we stained the harvested cells with β-galactosidase staining kit (G8861, Solarbio). SA-β-Gal staining kit is a kit for staining a...

Embodiment 2

[0043] Place C6 cells at 37°C, 5% CO 2 cultured in an incubator. The medium is DMEM + 10% FBS + 1% PS. After the cells are congested, use 1×10 5 The density was inoculated, and when the cells grew to about 70%, they were treated with 20 g / L D-galactose. Change the medium every other day to prevent D-galactose from becoming invalid. On days 2, 4, 6, and 8, the cells growing on the glass slides were harvested respectively. When the drug treatment time exceeds 3 days, subculture is carried out, a part of the cells are transferred to the slide, and another part of the cells are transferred to the culture dish without the slide. And so on until the 8th day sample is collected.

[0044] We stained the harvested cells with β-galactosidase staining kit. For cultured cells, remove the medium and wash the plate 3 times with HBSS; 2) Add 1 mL of β-galactosidase fixation solution and fix the cells at room temperature for 10 min; 3) Remove the cell fixation solution; wash the plate w...

Embodiment 3

[0046] The difference between embodiment 3 and embodiment 1 is that the treatment concentration of D-galactose is 5g / L, which does not change with the parameters.

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Abstract

The invention relates to a disease model construction method and in particular relates to a method for preparing a glioma cell senescence model by using D-galactose. The method comprises the followingsteps: (1) culturing glioma cells in an incubator; (2) after culture is completed, adding D-galactose into a cell culture medium, and performing treatment for a certain period of time; and (3) afterD-galactose treatment is completed, performing dyeing by using a dyeing liquid containing beta-galactosidase, microscopically observing image records, and calculating the SA-beta-Gal dyeing positive rate. The method overcomes defects that in the prior art, a method for preparing a senescence model generally consumes great time and labor, the cost is high, in addition, the success rate is low, meanwhile, reagents used in the method have great toxicity, and mass mortality of cells is frequently caused, thus has the advantages of being mild in treatment condition, short in time and easy to operate, in addition, high in success rate and good in effect, and provides both novel ideas for treatment on glioma and model bases for development of antitumor medicines.

Description

technical field [0001] The invention relates to a method for establishing a disease model, in particular to a method for preparing a senescence model of neuroglioma cells by using D-galactose. Background technique [0002] Glioma, referred to as glioma or glioma, is the most common primary central nervous system tumor, accounting for about 40% to 50% of all primary intracranial tumors (Kalpathy-Cramer et al., 2014 ; Linz, 2010). Gliomas can be divided into astrocytomas, oligodendrocytomas, ependymomas and Mixed glioma. The World Health Organization classifies gliomas into grades one to four. The first-grade glioma can be cured by surgical resection; the second-grade glioma can be treated with comprehensive treatment such as surgery, radiotherapy, and chemotherapy, and the survival period may reach five to ten years; The normal period will not exceed two to five years. Finding an effective treatment method to improve the therapeutic effect of malignant glioma has become ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09G01N1/30
CPCC12N5/0693G01N1/30G01N2001/302
Inventor 黄智慧徐星星沈细亚谢恬王莹冯文金
Owner HANGZHOU NORMAL UNIVERSITY
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