Human brain glioma cell line, and establishing method and application thereof

A technology of human glioma and glioma, which is applied in the field of cell lines, can solve the problems of unsatisfactory treatment effects of brain tumors, and achieve the effect of enriching the world's brain tumor cell bank and stabilizing its properties

Active Publication Date: 2014-03-12
SHANGHAI CHEMPARTNER CO LTD +1
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  • Abstract
  • Description
  • Claims
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  • Human brain glioma cell line, and establishing method and application thereof
  • Human brain glioma cell line, and establishing method and application thereof
  • Human brain glioma cell line, and establishing method and application thereof

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Embodiment 1

[0043] The preparation of embodiment 1GLC-001 cell

[0044] A fresh clinical glioma resection specimen (male, 52 years old, glioblastoma multiforme) was obtained from Nantong Cancer Hospital, and immediately immersed in pre-cooled sterile HBSS buffer (containing 500 U / ml penicillin G, 500 μg / ml streptomycin sulfate and 1.25 μg / ml amphotericin B). Primary culture: In a biological safety cabinet, wash the specimen with fresh sterile HBSS buffer, cut it into small pieces of 20-30 cubic millimeters, use 20ml of phosphate buffer solution containing 0.2% type II collagenase to digest the tumor piece, and put Dispersed cells were resuspended in 5ml DMEM / F12 complete culture medium, the composition of which said DMEM / F12 complete culture medium included: 1×B27 nutritional factor (StemCell Technology, 07153), 20ng / ml human epidermal growth factor, 20ng / ml human Fibroblast growth factor, 100 U / ml penicillin G and 100 μg / ml streptomycin sulfate were placed in cell culture flasks and pl...

Embodiment 2

[0048] The biological characteristics and application of embodiment 2GLC-001 cells

[0049] The present invention adopts DMEM / F12 complete culture medium to culture and purify GLC-001 cells, and then uses RPMI 1640 culture medium containing fetal bovine serum to cultivate them, so that they can grow long-term in vitro and pass stably. When the cells are passed to more than 20 generations, the cell properties are gradually stabilized, and the relevant biology, genetics and tissue source identification are carried out until the 50th generation has the same stable properties. Through experimental observation and verification, the main body of GLC-001 grown in vitro is spindle-shaped, with irregular branches or bifurcations. When the cell density is high, it loses contact growth inhibition, and grows in a cross-stacking state, and grows outward from the aggregation. Genetic studies have confirmed that the cells are aneuploid, with severe chromosome number and structural aberration...

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Abstract

The invention provides a human brain glioma cell line and an establishing method and application thereof. The human brain glioma cell line is preserved in China Center for Typical Culture Collection with an accession number of CCTCC No. C201115. The human brain glioma cell line is established by subjecting brain glioma cells originated from a clinic specimen to primary culture and subculture, can be directly used for in vitro drug screening or generation of human brain glioma in mammals and is used for establishing a human brain glioma animal model and screening candidate drugs used for treating human brain glioma. The human brain glioma cell line has stable properties, can realize stable multiple passages and maintain stable properties even after in vitro passage to the 50th generation; pathogenesis of human brain glioma, drug susceptibility, transitivity and other related characteristics can be analyzed in vitro and in vivo, so two correlated in-vitro and in-vivo anti-brain glioma drug screening platforms can be established, and novel experimental materials closer to clinic oncobiological characteristics are provided for research on human brain glioma.

Description

technical field [0001] The invention belongs to the field of cell lines, in particular to a human glioma cell line and its establishment method and application. Background technique [0002] Tumors that grow in the brain are collectively called brain tumors, including primary brain tumors that originate from the brain parenchyma and secondary brain tumors that metastasize to the brain from other parts of the body. Primary brain tumors are divided into benign and malignant according to their biological characteristics. Benign brain tumors grow slowly, have relatively complete capsules, do not infiltrate surrounding tissues, and are well differentiated; malignant brain tumors grow rapidly, have no capsule, have indistinct boundaries, show infiltrative growth, and are poorly differentiated. [0003] Among brain tumors, glioblastoma has the highest incidence rate, accounting for about 40.49%, and the peak age of onset is between 30 and 40 years old, or 10 to 20 years old. Glio...

Claims

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Application Information

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IPC IPC(8): C12N5/09A01K67/027C12Q1/02A61K49/00
Inventor 秦宵然王科张一心谢付波王建红闻丹忆季从飞赵奇红
Owner SHANGHAI CHEMPARTNER CO LTD
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