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Preparation method and application of a BCSC (breast cancer stem cell) antigen

A technology of breast cancer stem cells and antigens, applied in the fields of bioengineering and biomedicine, can solve the problem of chemotherapy drug insensitivity and achieve the effect of improving tumor killing ability

Inactive Publication Date: 2018-12-28
上海莱馥生命科学技术有限公司
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  • Abstract
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  • Claims
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Problems solved by technology

The proportion of CSC in tumor tissue is very low, but they are mainly in the quiescent phase and are not sensitive to chemotherapy drugs, so CSC may be the root cause of tumor recurrence after treatment

Method used

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  • Preparation method and application of a BCSC (breast cancer stem cell) antigen
  • Preparation method and application of a BCSC (breast cancer stem cell) antigen
  • Preparation method and application of a BCSC (breast cancer stem cell) antigen

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preparation example Construction

[0031] The invention discloses a preparation method of breast cancer stem cell antigen, comprising the following steps:

[0032] (1) Culturing breast cancer stem cells;

[0033] (2) adding hypochlorous acid into the solution for cultivating breast cancer stem cells, mixing evenly, and inducing apoptosis of breast cancer stem cells;

[0034] (3) The breast cancer stem cells treated with hypochlorous acid were lysed, and the cell lysate was collected to obtain the antigen.

Embodiment 1

[0036] Breast cancer stem cell medium is DMEM / F12 medium, and added: 20ng / mL EGF, 20ng / mL bFGF, 20ng / mL PDGF-AA and 1×B27. DMEM / F12 medium was purchased from Thermo Fisher; EGF was epidermal growth factor, purchased from PeproTech; bFGF was basic fibroblast growth factor, purchased from PeproTech; PDGF-AA was a platelet-derived factor; B27 is a stem cell growth factor purchased from Thermo Fisher.

[0037] Proceed as follows:

[0038] 1. Dissociate fresh breast cancer tumor tissue and isolate individual tumor cells

[0039] 1.1 Transfer the fresh breast cancer tissue block to a petri dish with sterile tweezers, wash the tissue block twice with normal saline, and discard the waste liquid. A small amount of serum-free 1640 medium was dropped on the tissue block, and the tissue block was cut into pieces with scissors. Transfer the shredded tissue pieces to a 50mL centrifuge tube and centrifuge at 1500rpm for 10 minutes; absorb the waste liquid, and add digestive solution (5mg / ...

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Abstract

The invention discloses a preparation method and an application of a BCSC (breast cancer stem cell) antigen. The method comprises the following steps: culturing a BCSC, adding hypochlorous acid to a solution for culturing the BCSC for uniform mixing, inducing apoptosis of the BCSC, cracking the BCSC treated by hypochlorous acid, and collecting a cell lysate to obtain the antigen, wherein hypochlorous acid added to the solution for culturing the BCSC has concentration of 20-80 mu M and treatment time of hypochlorous acid is 30-60 min. The antigen prepared with the method can be used for preparing tumor DC vaccine. The method for collecting the antigen after treating the BCSC with hypochlorous acid is not only economical and simple, but also can more strongly activate immune response of antigen-specific T cells and improve tumor killing capacity of the T cells.

Description

technical field [0001] The invention relates to the fields of bioengineering and biomedicine, in particular to a preparation method and application of a breast cancer stem cell antigen. Background technique [0002] Breast cancer is a malignant tumor that occurs in the epithelial tissue of the breast. 99% of breast cancer occurs in women, and only 1% in men. Since the 1990s, the incidence of breast cancer in China has grown twice as fast as the global average, especially in urban areas. At present, breast cancer is the cancer with the highest incidence rate in Chinese women, ranking fifth in the death of malignant tumors in women. For breast cancer, there are about 25 countries and regions with a 5-year survival rate greater than 85%, among which 3 Asian countries include Israel, South Korea and Japan. The 5-year survival rate of breast cancer in my country is in the middle of Asian countries and regions, lower than that of Israel, South Korea and Japan, and slightly lower...

Claims

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Application Information

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IPC IPC(8): C07K14/47C12N5/095A61K39/00A61P35/00
CPCA61K39/0011A61K2039/5154A61P35/00C07K14/4748C12N5/0695
Inventor 李新峰
Owner 上海莱馥生命科学技术有限公司
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