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3D culture medium for brain glioma organs and application thereof

A technology of glioma and organoids, applied in the field of biomedicine, can solve the problems of colon, stomach, prostate and pancreas and other tissues that have been successfully constructed in vitro in organ culture, so as to avoid hypoxic damage and promote cluster growth , the effect of accelerating the proliferation speed

Active Publication Date: 2021-02-02
ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, although organoids from tissues such as colon, stomach, prostate, and pancreas have been successfully constructed in vitro using organoid culture, there are few reports on the use of surgically resected glioma tissue for organoid culture in glioma

Method used

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  • 3D culture medium for brain glioma organs and application thereof
  • 3D culture medium for brain glioma organs and application thereof
  • 3D culture medium for brain glioma organs and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0047] This embodiment provides a glioma organoid medium, including Neurocult NS-A basal medium and specific additive factors; the specific additive factors include components at the following final concentrations: B27 additive, 2%; Thiazovivin L-Glutamine, 0.1%; HGF, 25ng / ml; Insulin, 0.05%; FGF-2, 30ng / ml; dorsomorphin, 2.5μM; SB-431542, 25μM; EGF, 250ng / ml; BSA 5%; HEPES, 20mM; Heparin, 0.05%; human NLGN3, 100nM; double antibody, 1%; the aforementioned percentage concentration represents the mass concentration. The preparation method of the culture medium is as follows: according to the above-mentioned composition and ingredients of the culture medium, specific additive factors are added to the base culture medium and mixed evenly.

Embodiment 2

[0049] This embodiment provides a glioma organoid medium, including Neurocult NS-A basal medium and specific additive factors; the specific additive factors include components at the following final concentrations: B27 additive, 1%; Thiazovivin , 10μM; L-Glutamine, 0.05%; HGF, 50ng / ml; Insulin, 0.06%; FGF-2, 10ng / ml; dorsomorphin, 3μM; SB-431542, 20μM; EGF, 500ng / ml; 2%; Heparin, 0.01%; human NLGN3, 200nM; double antibody, 0.5%; the aforementioned percentage concentration represents the mass concentration. The preparation method of the culture medium is the same as in Example 1.

Embodiment 3

[0051] This embodiment provides a glioma organoid medium, including Neurocult NS-A basal medium and specific additive factors; the specific additive factors include components at the following final concentrations: B27 additive, 5%; Thiazovivin , 1μM; L-Glutamine, 0.2%; HGF, 10ng / ml; Insulin, 0.03%; FGF-2, 50ng / ml; dorsomorphin, 2μM; SB-431542, 30μM; 9%; Heparin, 0.1%; human NLGN3, 50nM; double antibody, 5%; the aforementioned percentage concentration represents the mass concentration. The preparation method of the culture medium is the same as in Example 1.

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Abstract

The invention provides a 3D culture medium for brain glioma organs and application thereof, the culture medium comprises a basic culture medium and specific additive factors, and the specific additivefactors comprise the following components with a final concentration: 1-5% of B27 additive; 1-10 [mu]M of Thiazovivin; 0.05-0.2% of LGlutamine; 10-50 ng / ml of HGF; 0.03-0.06% of Insulin; 10-50 ng / mlof FGF-2; 2-3 [mu]M of domomorphin ; 20-30 [mu]M of SB-431542; 100-500 ng / ml of EGF; 10-30 mM of HEPES; 1%-10% of BSA (Bovine Serum Albumin); 0.01%-0.1% of Heparin; 50-200 nM of human source NLGN3; 0.5-5% of a double antibody; wherein the percentage concentration is the mass concentration. The culture medium of the invention is used for culturing brain glioma organs, so that brain glioma tissue cells can better show the inherent activity characteristics in the culture process, which are highly similar to the comprehensive characteristics of living brain tissues.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a 3D medium for glioma organoids and an application thereof. Background technique [0002] Glioma is a common malignant tumor of the central nervous system, accounting for about 40%-50% of intracranial tumors, mostly in the form of infiltrative growth, with unclear boundaries with the surrounding brain tissue, and difficult to be completely removed by surgery. It is easy to relapse after surgery; generally speaking, the higher the grade of the tumor, the higher its malignancy, and the faster the recurrence after surgery. At present, the treatment of glioma is mainly based on surgical resection combined with postoperative radiotherapy and chemotherapy, but the prognosis is poor. In recent years, no breakthrough has been made in the treatment of glioma. [0003] However, currently, there are only a handful of in vitro glioma models used in drug development and bas...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09C12N5/079
CPCC12N5/0622C12N5/0693C12N2501/11C12N2501/115C12N2501/12C12N2501/13C12N2501/15C12N2501/33C12N2501/405C12N2501/73C12N2513/00
Inventor 陈泽新于言徐丛黄敏
Owner ACCURATE INT BIOTECHNOLOGY (GUANGZHOU) CO LTD
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