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
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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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