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Human glioma cell line and establishment method and applications thereof

A technology of human glioma and its establishment method, which is applied in the field of human glioma cell lines and its establishment, can solve the problems of restricting preclinical research and scarcity of glioma cell lines, and achieve strong proliferation ability and original Easier to succeed in generation training, and easier to establish a line

Active Publication Date: 2019-04-12
BEIJING NEUROSURGICAL INST
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, at present, there are only a handful of glioma cell lines used in drug development and basic research in the world, such as LN-229, U-87MG, U-118MG and U-138MG cell lines are all derived from Caucasian Humans, and glioma cell lines with Asian genetic background are even rarer. Compared with the large number of glioma patients in clinical practice, the scarcity of glioma cell lines seriously restricts preclinical research

Method used

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  • Human glioma cell line and establishment method and applications thereof
  • Human glioma cell line and establishment method and applications thereof
  • Human glioma cell line and establishment method and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0046] Example 1 Construction method of human glioma cell line BT-127

[0047] 1. Patient tissue sample processing

[0048] 1.1 Overview of the patient's condition:

[0049] The patient, male, 37 years old, was diagnosed with glioblastoma. He had undergone glioma surgery twice before the resection of glioma tissue. The lesion also metastasized under the scalp. It can be seen that the invasiveness of this glioma is very strong. There are few cases of glioma metastasizing to the extracranial. The number of cultured glioma cell lines is even less, so the human brain glioma cell line of the present invention has important significance for the study of glioma invasion and metastasis and drug testing.

[0050] In addition, the patient underwent radiotherapy and chemotherapy before the resection of glioma tissue, so the human glioma cell line of the present invention is very likely to be resistant to radiotherapy and chemotherapy, and the PDX model thus established may have the abi...

Embodiment 2

[0065] Morphological observation of embodiment two BT-127 cells

[0066] Place the culture flask for cultivating BT-127 cells under an inverted microscope and observe under bright field. figure 1 It is a photo of BT-127 cells cultured in G1 generation. The results show that the main body of BT-127 cells is spindle-shaped, and there are irregular branches or bifurcations ( figure 1 -A, 100×); when the cell density was high, the contact growth inhibition was not obvious, and it was in a state of cross-stacking growth, and radiated outward from the aggregation ( figure 1 -B, 100×).

[0067] figure 1 -C is a photo of BT-127 cells cultured in G20 generation. figure 1 -D is the photo of BT-127 cells G50 generation culture. Depend on figure 1Comparison of the four pictures A-D shows that after 50 subcultures, the morphology of the cells remains unchanged, which shows that the BT-127 cells prepared by the present invention can be stably subcultured, which is helpful for long-term...

Embodiment 3

[0068] Karyotype analysis of embodiment three BT-127 cells

[0069] The BT-127 cells growing in the logarithmic phase were taken, and colchicine was added to make the final concentration 0.2 μg / ml, and cultured in a 37° C. incubator for 4 hours. Cells in the mid-division stage were collected and fixed with a fixative solution (methanol: glacial acetic acid = 3:1 (V / V), ready-to-use) for about 1 hour, and then the cell suspension was dropped on a pre-cooled slide, and used Stained with Giemas staining solution, and the number of chromosomes was counted under a microscope. see results figure 2 , after the BT-127 cells were continuously passaged, the chromosomes still maintained the characteristics of the chromosomes of human-derived tumor cells, and the modal number (M) of the chromosomes was concentrated between 45 and 55, accounting for 70%. It can be seen that the BT-127 cells are aneuploid, Chromosome number and structural aberrations are serious, consistent with the gene...

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Abstract

The invention discloses a human glioma cell line. The preservation number of the human glioma cell line in the China General Microbiological Culture Collection Center is CGMCC NO.16693, and the preservation date is: November 07, 2018. The human glioma cell line of the invention can be stably multi-passaged, has tumorigenicity, can be prepare into a tumor animal model, and can be used for researchand development of human glioma diagnostic reagents, screening of human glioma drugs, and in vitro-studying of human gliomas. Moreover, the present invention differentiates glioma stem cells into glioma cells so as to firstly reduce the number of hetero cells, and after the glioma stem cells are differentiated into the glioma cells, the growth rate is fast, the proliferative ability is strong, andthe characteristics of pathological tissues of brain glioma is retained.

Description

technical field [0001] The invention belongs to the field of medicine, and in particular relates to a human brain glioma cell line and its establishment method and application. Background technique [0002] Glioma is a common tumor of the central nervous system, with an incidence of about 3-5 / 100,000 people. Histology is divided into four grades: grade I and grade II are low malignancy and have benign biological characteristics; grades III and IV are high-grade gliomas, especially glioblastoma, and the median survival time is only 12-18 The median survival period of anaplastic glioma (WHO grade III) is 2-5 years, and the median survival period of low-grade glioma (WHO grade I, II) is only 4-10 years. [0003] Although neurosurgery skills are constantly improving, surgical instruments are becoming more and more advanced, and new progress has been made in combination with radiotherapy and chemotherapy. In addition, there are some new treatment methods, such as immunotherapy, ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/09A01K67/027C12Q1/02A61K49/00C12R1/91
Inventor 张俊文刘福生方胜宋文杰
Owner BEIJING NEUROSURGICAL INST
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