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Preparation method of model used for in vitro glioma stem cell drug screening

A glioma stem cell and application model technology is applied in the field of preparation of an in vitro glioma stem cell drug screening application model.

Inactive Publication Date: 2017-05-03
袁国红
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] Glioma is the most common tumor of the central nervous system. High-grade glioma has a strong ability to invade. Therefore, the effect of surgery, prevention, chemotherapy and other treatment methods is not very good. It tends to recur and has a poor prognosis. serious threat to people's health
However, studies have found that not all glioma cells can proliferate indefinitely, only brain glioma stem cells have the ability of unlimited proliferation and self-renewal. Significance of recurrence, mechanism of invasion, and effective treatment

Method used

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Embodiment Construction

[0008] In order to further illustrate the present invention, the primary culture of glioma cells from patients is taken as an example to further describe the present invention in detail.

[0009] The preparation method that can be used for the application model of glioma stem cell drug screening in vitro: including glioma stem cell isolation, glioma stem cell suspension culture and glioma stem cell adhesion culture.

[0010] The first step is to isolate glioma stem cells, and the specific steps include: (1) Put the glioma tissue freshly obtained from the patient's brain in the neural stem cell basal medium containing 15% PS, and transport it on ice; (2) Take out the tissue , placed in neural stem cell basal medium containing 15% PS, rinsed twice, and cut off debris such as blood vessels with microsurgical scissors; (3) Microsurgical scissors cut up the tissue in a 10cm culture dish, and added 5ml of pre-warmed Accumax digestive enzyme, digested at 37°C for 30 minutes, pipettin...

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Abstract

The invention belongs to the technical field of genetic engineering, and more specifically relates to a preparation method of a model used for in vitro glioma stem cell drug screening. The preparation method comprises steps of glioma stem cell separation, glioma stem cell suspension culturing, and glioma stem cell adhesion culturing. The step of glioma stem cell separation comprises following specific steps: 1, glioma tissue is acquired; 2, the glioma tissue is collected, and is subjected to rinsing; 3, the glioma tissue is cut into pieces, digestive enzyme is added, digestion is carried out for 30min at 37 DEG C, and dissociating is carried out every 5min; 4, reaction is stopped, and centrifugation is carried out; 5, washing is carried out; 6, counting, inoculating, and culturing are carried out. The step of glioma stem cell adhesion culturing and freeze preservation comprises following specific steps: 1, after aggregation of cells obtained in the former step, digestion is carried out so as to obtain single cells, and centrifugation is carried out; 2, counting and subcultring are carried out, culturing solution is changed in the next day of subcultring, and then is changed every other day; 3, when density is larger than 90%, digestion into single cells is carried out, and subcultring or freeze preservation is carried out. The primary culturing method of glioma cells is simple, and is convenient in operation, and adherent primary culturing success rate of glioma stem cells is high.

Description

technical field [0001] The invention relates to the technical field of genetic engineering, in particular to a preparation method for an application model of glioma stem cell drug screening in vitro. Background technique [0002] Glioma is the most common tumor of the central nervous system. High-grade glioma has a strong ability to invade. Therefore, treatment methods such as surgery, prevention, and chemotherapy are not effective, tend to recur, and have a poor prognosis. serious threat to people's health. However, studies have found that not all glioma cells can proliferate indefinitely, only brain glioma stem cells have the ability to proliferate indefinitely and self-renew. Mechanisms of recurrence, invasion, and effective treatment are of great interest. Contents of the invention [0003] The purpose of the present invention is to provide a preparation method that can be used for the application model of glioma stem cell drug screening in vitro, with simple steps, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/095
CPCC12N5/0695
Inventor 袁国红赵曙红刘彬刘庆柱杨亮
Owner 袁国红