Extraction method and application of tumor exosome vaccine

A tumor exosome and extraction method technology, which is applied in the field of tumor exosome (exosome) vaccine extraction, can solve the problem of inability to extract miRNA antigens, and achieve the goal of improving tumor-specific immunogenicity and enhancing killing ability. Effect

Inactive Publication Date: 2013-10-09
ZHEJIANG UNIV
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Problems solved by technology

The original method of extracting exosome only extracts the complete vesicle structure of exosome, but a large number of miRNA and tumor-associated antigens in the vesicle cannot be extracted

Method used

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  • Extraction method and application of tumor exosome vaccine
  • Extraction method and application of tumor exosome vaccine
  • Extraction method and application of tumor exosome vaccine

Examples

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Embodiment 1

[0016] Application of exosome in immunotherapy of cholangiocarcinoma cells

[0017] 1. Extract exosome: Fetal bovine serum was centrifuged at 100,000×g for 6 hours at 4°C to remove endogenous exosome. Human cholangiocarcinoma cells (RBE cells) were cultured in 1640 culture medium containing 10% fetal bovine serum for about After 72 hours, the medium supernatant was collected. Extract the exosome according to the Good Manufacturing Practice (GMP) standard: centrifuge the supernatant at 300×g for 5 minutes, transfer to a new centrifuge tube and centrifuge at 800×g for 30 minutes, transfer to a new centrifuge tube and centrifuge at 10,000×g for 30 minutes to remove cell debris; The supernatant was then centrifuged at 1,500×g for 30 minutes with a 100 kd ultrafiltration tube from Millipore Company; the concentrated solution was transferred to a 30 g / L sucrose heavy water pad, and ultracentrifuged at 100,000×g for 60 minutes. Take out the sucrose heavy water pad, dilute it into 50...

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Abstract

The invention provides an extraction method and application of a tumor exosome vaccine. According to the invention, exosome is successfully extracted from the supernatant of a nutrient solution of human bile duct cancer cells, and an exosome lysis ultrafiltrate not containing miRNA is extracted by using an ultrafiltration lysis method. The exosome lysis ultrafiltrate not containing miRNA obtained in the invention can obviously stimulate immune cells to generate a specific tumor killing effect, greatly improve the killing effect, i.e., specific immune response, of the immune cells on a tumor, reduce the level of immune escape of tumor cells and enhance killing capability of the human immune cells on the tumor cells.

Description

technical field [0001] The invention belongs to the field of biological technology, and relates to a method for extracting a human tumor vaccine, in particular to an extraction method and application of a tumor exosome vaccine. Background technique [0002] Surgical treatment is a routine treatment for tumors. In addition to surgical treatment, adoptive cellular immunotherapy is currently a very promising tumor treatment method. Compared with other treatment methods, adoptive cellular immunotherapy has the advantages of high specificity, less side effects, improvement of clinical signs, and prolongation of patient survival time. In vitro and animal experiments have shown that adoptive cells have a significant specific killing effect on tumor cells. [0003] The exosome is a small vesicle with a diameter of 50-100nm secreted by the fusion of the multivesicular bodies (MVBs) of many living cells with the cell membrane. Tumor cell-derived exosomes have been proven to express ...

Claims

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Application Information

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IPC IPC(8): A61K35/16A61P35/00
Inventor 曹利平陈炯煌阙日升
Owner ZHEJIANG UNIV
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