Human benign phyllodes tumor cell line GLK-1010 and applications thereof
A technology for tumor cells and benign tumors, applied in the field of tumor cells, can solve problems such as limited value, and achieve the effect of short establishment time and stable biological heredity
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Embodiment 1
[0018] The human phyllodes tumor cell line GLK-1010 of the present invention is obtained by the following method:
[0019] (1) Specimen collection and storage methods: After the complete mammary gland phyllodes tumor has been completely removed, the tumor tissue is cut in the middle with a sterile scalpel, and the vigorously proliferating tissue is taken in DMEM medium. The collection of specimens is carried out under the guidance of the surgeon and the pathologist to prevent the impact on the diagnosis of the pathology report; when the specimens are not used for follow-up operations, they should be stored in DMEM medium at 4°C, and the specimens should be used within 24 hours as much as possible .
[0020] (2) Primary culture: the tissue was washed 3 times with PBS, cut mechanically until it was as small as possible, digested with type III collagenase (1mg / ml, Worthington) and added to a 50ml centrifuge tube containing DMEM / F12, protected from light with tinfoil, 37°C, 180 r...
Embodiment 2
[0035] Embodiment 2 detects the expression situation of virus
[0036] Q-PCR (quantitative real-time polymerase chain reaction, referred to as quantitative PCR) was used to detect the expression level of the immortalized gene (HPV) introduced into the cell line GLK-1010, wherein the primer sequences used to amplify the corresponding gene were listed in Table 1 below. Show.
[0037] Table 1 Primer base sequence
[0038] Forward (forward primer) Reverse (reverse primer) GAPDH AGAGCCTCGAGGAGAAGTTCC (SEQ ID NO. 1) ACTAGGGAGTCAAGGACGGG (SEQ ID NO. 2) HPV TAACTTCTGTTCGGGGAGGC (SEQ ID NO. 3) AGGGAGGGAAAAGATCCCGA (SEQ ID NO. 4)
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