A kind of culture medium and culture method for culturing brain tumor cells in vitro

An in vitro culture and brain tumor technology, applied in the direction of tumor/cancer cells, culture process, tissue culture, etc., can solve the problems of drug screening for medulloblastoma

Active Publication Date: 2020-10-16
苏州麦迪耐斯医药科技有限公司
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Problems solved by technology

However, studies have shown that the above-mentioned tumor cells cultured by traditional adherent culture methods gradually inactivate the Shh pathway in vitro, making the tumor cells gradually stop proliferating, which brings great benefits to the basic research of medulloblastoma and corresponding drug screening. Difficulties

Method used

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  • A kind of culture medium and culture method for culturing brain tumor cells in vitro
  • A kind of culture medium and culture method for culturing brain tumor cells in vitro
  • A kind of culture medium and culture method for culturing brain tumor cells in vitro

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Embodiment 1

[0048] Experimental animals: conditional knockout transgenic mice Ptch1fl / fl Mice, Math1-Cre Mice, CB17 / SCIDMice, purchased from Jackson Laboratory, all experimental animals are in line with SPF level (Specific Pathogen Free, no specific pathogens), all animals are treated All operations have been legally and effectively approved; immunodeficiency mice (CB17 / SCID Mice), intracranial injection transplantation using a mouse brain stereotaxic instrument (KOPF, Nikon SMZ1000) and a 5ul micro-injector (Shanghai Gaoge).

[0049] Cell culture:

[0050] (1) The primary medulloblastoma cells were all extracted from the cerebellum of the above-mentioned conditional knockout adult mice, and the mouse cerebellar medulloblastoma tissue was digested with digestive juice, and the digestive juice components (10ml): 100U of papain ( Papain, Worthington Biochemical, LS003126), 2 mg of cysteine ​​(L-cysteine, Sigma) and 2500 U of DNase (DNase, Sigma, D4527), digested in a water bath at 37°C for ...

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Abstract

The invention provides a culture solution for culturing brain tumor cells in vitro. The culture solution comprises a nerve cell basal medium, and also comprises, based on the mass of the nerve cell basal medium, 0.5%-5% of a B27 trophic factor, 0.5%-5% of double antibodies- penicillin / streptomycin, 0.5%-5% of glutamine, 0.5%-5% of sodium pyruvate, 0.5%-1.5% of fetal bovine serum, and 2.5- 3.5 microgram / mL of recombinant Shh proteins. Cells obtained by applying the culture solution can proliferate and produce offspring, the Shh signaling pathway is in an activated expression state, the tumorigenicity is not affected, and the cells can withstand cryopreservation of 80 DEG C and achieve adherent differentiation, and thus a unique cell model is provided for studying the molecular mechanism ofmedulloblastoma development and the molecular mechanism of Shh signaling pathway regulation and an important tool is provided for high-throughput screening of chemotherapy drugs.

Description

technical field [0001] The invention belongs to the technical field of tissue cell culture, and in particular relates to a culture medium and a culture method for culturing brain tumor cells in vitro. Background technique [0002] Medulloblastoma (MB) is the most common malignant brain tumor in children. It mainly occurs in the cerebellum and can rapidly spread to the central nervous system. It is generally believed that MB includes at least 4 subtypes, namely Wnt subtype, Shh subtype, third subtype and fourth subtype. Among the four subtypes, the Shh subtype accounts for about 30% of human medulloblastoma, which is caused by abnormal activation of the Shh signaling pathway. [0003] Astrocytes are the most widely distributed type of glial cells in the mammalian brain and play a supporting role in the development and function of neurons. In recent years, the role of astrocytes in brain tumors has also been elucidated to some extent: it has been reported that astrocytes sec...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09A01N1/02
CPCA01N1/0221C12N5/0693C12N2500/30C12N2500/32C12N2501/41
Inventor 杨增杰程艳梁玉刚
Owner 苏州麦迪耐斯医药科技有限公司
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