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Preparation method of heat shock apoptosis glioma-loaded dendritic cell tumor vaccine

A technology for dendritic cells and gliomas, which can be used in antitumor drugs, medical preparations containing active ingredients, pharmaceutical formulations, etc., and can solve the problems of difficult to cure brain malignant gliomas, difficult to completely remove by surgery, etc.

Inactive Publication Date: 2012-07-11
江苏迈健生物科技发展股份有限公司
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Problems solved by technology

Due to the rapid growth and infiltrative growth along the nerve fibers, it is difficult to completely remove it by surgery; at the same time, it is only moderately sensitive to chemotherapy and radiotherapy
Therefore, the current internationally recognized sequential treatment plan of conventional surgery, radiotherapy, and chemotherapy is difficult to cure malignant glioma of the brain.

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  • Preparation method of heat shock apoptosis glioma-loaded dendritic cell tumor vaccine
  • Preparation method of heat shock apoptosis glioma-loaded dendritic cell tumor vaccine

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Embodiment Construction

[0013] like figure 1 The schematic flow chart of the steps of the preparation method of the heat-shocked apoptotic glioma-loaded dendritic cell tumor vaccine of the present invention is shown, and the specific steps of the embodiment are as follows:

[0014] S1. Preparation and detection of heat shock apoptotic glioma cells:

[0015] The human glioma cell line U251 was introduced from ATCC in the United States, and subcultured according to conventional methods. Adjust the cell concentration to 10 with RPMI1640 medium 6 / ml, put the cells at 40°C, 5% CO 2 After 3 hours in the incubator, take out the cell culture bottle, add betulinic acid to a final concentration of 10 μg / ml, and move the cell culture bottle to 37°C, 5% CO 2 Continue culturing in the incubator to induce apoptosis. Apoptotic cells were harvested, washed 4 times with sterile PBS, and apoptotic tumor cell antigens were obtained.

[0016] After the tumor cells were treated at 40°C for 3 hours, the cells were d...

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Abstract

The invention provides a preparation method of a heat shock apoptosis glioma-loaded dendritic cell tumor vaccine. The method comprises the following steps of: setting a tumor cell in a heat shock state; inducing a cell high-expression heat shock protein (HSP); inducing cell apoptosis by using a chemical medicament to obtain an apoptosis cell antigen which is rich in HSPs; and culturing the prepared antigen with an immature dendritic cell (DC) in a hybrid way to obtain a mature DC vaccine. A heat shock glioma cell-loaded DC vaccine has high apoptosis rate, and has the phenotypic traits of a mature DC as proved by flow cytometry detection, the average value of universally-recognized marks HLA-DR<+>CD11C<+> of the DC is 90.90 percent, the average value of mature marks CD11c<+>CD86<+> is 73.03 percent, the average value of CD11c<+>CD83<+> is 86.04 percent, and CD83 and CD86 are expressed efficiently. The DC vaccine prepared by loading with a heat shock method has high maturity, and can be used for inducing immune response.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to a preparation method of a dendritic cell tumor vaccine loaded with heat shock apoptosis brain glioma. Background technique [0002] Brain malignant glioma is the most common primary malignant tumor of the central nervous system. Due to the rapid growth and infiltrative growth along the nerve fibers, complete surgical resection is difficult; meanwhile, it is only moderately sensitive to chemotherapy and radiotherapy. Therefore, the current internationally recognized sequential treatment plan of conventional surgery, radiotherapy, and chemotherapy is difficult to cure brain malignant glioma. Immunotherapy is to achieve the real purpose of cure by adjusting and improving the function of the body's immune system and eliminating residual tumor cells. Currently, DC-based cell vaccine therapy is an active immunotherapy and is becoming a research hotspot. DC is the most powerful antigen p...

Claims

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Application Information

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IPC IPC(8): A61K39/00A61P35/00
Inventor 陆华
Owner 江苏迈健生物科技发展股份有限公司
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