Drug-resistant glioblastoma cell line of primary temozolomide and bevacizumab, construction method and application of cell line

A technology for glioblastoma and bevacizumab, applied in the direction of microorganism-based methods, animal cells, tumor/cancer cells, etc., can solve problems such as not being able to guide the application well, and achieve stable and stable properties genetic effect

Inactive Publication Date: 2014-01-22
李文斌 +2
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] Most of the existing glioma cell lines at home and abroad do not have the characteristics of primary drug resistance, and some drug-resistant cell lines i

Method used

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  • Drug-resistant glioblastoma cell line of primary temozolomide and bevacizumab, construction method and application of cell line
  • Drug-resistant glioblastoma cell line of primary temozolomide and bevacizumab, construction method and application of cell line
  • Drug-resistant glioblastoma cell line of primary temozolomide and bevacizumab, construction method and application of cell line

Examples

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Embodiment 1

[0046] Example 1 Construction of primary drug-resistant glioblastoma cell lines

[0047] 1. Experimental method

[0048] Surgical specimens obtained from patients with human glioblastoma were surgically obtained, and the dispersed tumor cells were obtained by enzymatic digestion. They were placed in culture flasks containing medium, and continuously subcultured to 50 generations to obtain glioblastoma with stable passage in vitro. cell line. The specific implementation steps are as follows:

[0049] 1. Processing of Surgically Obtained Tumor Tissue

[0050] Put the tissue samples obtained from the operation into the culture medium containing penicillin and streptomycin, and place them in a liquid nitrogen tank; bring them into the laboratory for processing within 30 minutes; wash the tissue blocks repeatedly in the ultra-clean workbench to remove Blood vessels and blood clots; use ophthalmic scissors to cut the tissue block into a size of about 3-5mm 3 Small pieces of smal...

experiment example 1

[0059] Experimental example 1 Biological characteristics identification experiment of primary drug-resistant glioblastoma LC-11 cell line

[0060] 1. Morphological observation

[0061] Place the culture flask of cultured LC-11 cells under an inverted microscope and take pictures under bright field. The cells grow adherently. It can be seen that the tumor cells have various shapes. Most of the cells are spindle-shaped, and some are triangular and polygonal. Loss of contact inhibition between cells, showing malignant growth, with the characteristics of overlapping growth ( Figure 4 ).

[0062] 2. Karyotype analysis

[0063] After the cultured LC-11 cells were placed at 4°C for 12 hours, colchicine was added to make the final concentration 4 μg / ml, and then placed in a 37°C incubator for 10 hours. Cells in the mid-division stage were collected, fixed with a fixative, and then the cell suspension was lowered onto a pre-cooled slide, stained with Giemas staining solution, and t...

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Abstract

The invention discloses a drug-tolerant glioblastoma cell line of primary temozolomide and bevacizumab, a construction method and application of the cell line. The construction method comprises the following steps: obtaining tissue specimens of patients with drug-resistant glioblastoma cell of primary temozolomide and bevacizumab through clinical screening, primarily culturing tumor cells through an enzyme digestion method, and continuously extending generation to 50 generations to obtain a strain of drug-tolerant glioblastoma cell line of primary temozolomide and bevacizumab; the microbial preservation number of the drug-tolerant glioblastoma cell line of primary temozolomide and bevacizumab is CGMCC No.8205. The glioblastoma cell line, disclosed by the invention, is stable in characteristics, stably heritable and high in tolerance to temozolomide and bevacizumab; the cell generation cycle lasts about 48 hours; the glioblastoma cell line has strong invasiveness and tumorigenicity. The glioblastoma cell line constructed by the construction method has important application prospect in aspects of screening medicines for treating glioblastoma and researching nosogenesis of the glioblastoma, and the like.

Description

technical field [0001] The present invention relates to a glioblastoma cell line, in particular to a primary temozolomide- and bevacizumab-resistant glioblastoma cell line and its construction method. The present invention further relates to the glioblastoma cell line The application of tumor cell lines in the screening of candidate drugs for temozolomide and bevacizumab-resistant glioblastoma, preparation of human glioblastoma models, etc. belongs to the field of construction of glioblastoma cell lines. Background technique [0002] Glioma is the most common intracranial malignant tumor, accounting for about 35% to 61% of intracranial tumors. Because of its abnormal biological characteristics such as rich blood vessels, invasive growth, and lack of apoptosis, gliomas are highly invasive, and the existence of the blood-brain barrier has brought many difficulties to the treatment of gliomas. There are more than 15,000 new cases of malignant glioma in the United States each y...

Claims

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Application Information

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IPC IPC(8): C12N5/09C12Q1/02A01K67/027C12R1/91
Inventor 李文斌江涛康勋丁卫邱晓光白雪佳陈静石蕊
Owner 李文斌
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