Human breast cancer pleural effusion metastasis cell line and its application

A human breast cancer, cell line technology, applied in tumor/cancer cells, animal cells, vertebrate cells, etc.

Active Publication Date: 2021-10-08
ZHEJIANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the HER2 targeting drug Herceptin can prolong the HER2 + The survival time of breast cancer patients, but about 15% of early-stage patients and more than 70% of metastatic patients show resistance to the drug

Method used

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  • Human breast cancer pleural effusion metastasis cell line and its application
  • Human breast cancer pleural effusion metastasis cell line and its application
  • Human breast cancer pleural effusion metastasis cell line and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] From a 69-year-old female patient with invasive ductal carcinoma of breast cancer (primary tumor ER + , PR + , HER2 + ) of metastatic pleural fluid, centrifuged to obtain malignant cells, resuspended in phosphate buffer saline containing double antibody and centrifuged 3 times. Resuspend the pellet with DMEM medium containing 10% fetal bovine serum, spread it on a sterile 10cm petri dish, and place it in 37°C CO 2 Cultivate in an incubator, do not move the culture dish for at least 2 days, which is conducive to the adhesion of primary tumor cells; when the cells grow to 70-80% in the dish, transfer them to 6 10cm culture dishes, observe under the microscope every day, and wait for observation Under the microscope, when the cells are oval and sheet-like distributed cell clusters, scrape off the surrounding fibroblasts, wash 3 times with phosphate buffer saline, add new medium to continue the culture; when the cell clusters grow to be visible to the naked eye, trypsin ...

Embodiment 2

[0045] The subcultured human breast cancer pleural effusion metastatic cell line 4Z-B-1 was used to observe the growth of the cells under an optical microscope (ZEISS inverted microscope).

[0046] Microscopic examination: if figure 1 As shown in middle a, the cytological morphological picture shows that the cells have good refraction, the cells are in good growth state, the background is clear, and most of the cells are nearly round. figure 1 Middle a is the morphology of primary breast cancer cells at passage 50.

Embodiment 3

[0048] (1) Sample preparation: when preparing two-dimensional cell slide samples, 5×10 4 A cultured 4Z-B-1 cell was plated on a sterile cell slide, and placed in an incubator for 24 hours, waiting for the cells to fully adhere to the wall; when preparing a three-dimensional cell spheroid sample, 1×10 7 Spread the cultured 4Z-B-1 cells into a low-adsorption culture dish, collect the formed cell spheres in a 15ml centrifuge tube after 72 hours, and then wash and centrifuge again to collect;

[0049] (2) Fix: Add 2.5% glutaraldehyde fixative to the cells for 2 hours, wash 3 times with 0.1M phosphate buffer, add 1% glutaric acid fixative for 2 hours, wash 3 times with 0.1M phosphate buffer;

[0050] (3) Dehydration: 30% ethanol → → 50% ethanol → → 70% ethanol → → 90% ethanol → → 95% ethanol → → 100% ethanol (2 times) in a refrigerator at 4°C, each process is maintained for 15 -20min;

[0051] (4) Drying: place the sample at the critical point of the sample to dry;

[0052] (5) ...

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Abstract

The invention belongs to the field of microbial animal cell lines, and in particular relates to human breast cancer pleural effusion transfer cell lines and applications. The human breast cancer pleural effusion metastasis cell line includes: named human breast cancer pleural effusion metastasis cell line 4Z-B-1, and preserved in the China Center for Type Culture Collection, the preservation number is: CCTCC NO: C202123 human breast cancer pleural effusion metastasis cell line . The human breast cancer pleural effusion metastasis cell line 4Z-B-1 of the present invention is established from a Chinese source, and the establishment time is relatively short, and the biological genetic traits are stable. Taking the human breast cancer pleural effusion metastasis cell line as a research model, for Understanding Primary HER2 in the Chinese Population + The pathogenesis of breast cancer helps a lot.

Description

technical field [0001] The invention belongs to the field of microbial animal cell lines, and in particular relates to human breast cancer pleural effusion transfer cell lines and applications. Background technique [0002] Breast cancer is the most common malignancy in women worldwide. In my country, the incidence rate of breast cancer has gradually increased. In some coastal economically developed areas, the incidence rate of breast cancer has accounted for the first place in women's malignant tumors. Breast cancer is a highly heterogeneous malignant tumor. After decades of scientific research and technology development, it can be divided into five subtypes by using gene microarray technology: ductal type A, ductal type B, basal Cell-like, HER2-overexpressed and normal-breast-like types have significantly improved the therapeutic effect of clinical breast cancer patients and reduced the physical and mental pain suffered by patients during the treatment of the disease. ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09C12Q1/02A01K67/027C12R1/91
CPCA01K67/0271A01K2207/12A01K2227/105A01K2267/0331C12N5/0693C12N2503/02G01N33/5011G01N2500/10
Inventor 王林波李召卿周济春陈丽妮陈聪张迅陈永霞杨静静周雨露毛米莎许伶居思薇
Owner ZHEJIANG UNIV
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