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
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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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