Tumor cell apoptotic body and its preparation method and application

A tumor cell apoptosis and tumor cell technology, applied in the field of tumor cell apoptotic bodies and their preparation, can solve the problems of damage to normal cells, adverse reactions, poor targeting, etc., and achieve reduced treatment costs, good treatment effects, and production low cost effect

Active Publication Date: 2021-03-19
SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

Cancer treatment methods mainly include surgical treatment, radiotherapy and drug therapy. However, these traditional cancer treatment methods are poorly targeted and highly toxic. While killing tumor cells, they will also damage normal cells and cause adverse reactions. Therefore, there are certain limitations

Method used

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  • Tumor cell apoptotic body and its preparation method and application
  • Tumor cell apoptotic body and its preparation method and application
  • Tumor cell apoptotic body and its preparation method and application

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preparation example Construction

[0060]One embodiment of the present invention provides a method for preparing tumor cell apoptotic bodies, comprising the following steps: culturing tumor cells in a hypoxic environment to induce tumor cell apoptosis and generate apoptotic bodies.

[0061] The hypoxic environment refers to a state where cells cannot obtain oxygen normally, and the oxygen concentration is preferably not higher than 6%, more preferably 1%-6%. The hypoxic environment can also be formed by the following method: adding cobalt chloride, defaminated ferric and / or sodium sulfite and other compounds that induce cell hypoxia in the medium for culturing tumor cells, and the addition amount is added to can up-regulate the tumor cell hypoxia-inducible factor 1α (HIF-1α).

[0062] The apoptotic bodies of tumor cells with higher activity can be obtained by the above preparation method, and the apoptotic bodies can effectively stimulate the immune activity in tumor carriers, thereby having excellent anti-tumo...

Embodiment 1

[0076] Example 1 Preparation of Tumor Cell Apoptotic Body in Hypoxic Environment

[0077] 1. Induce tumor cell apoptosis

[0078] In an oxygen-deprived environment (O 2 The concentration is 1%-6%), continue to culture breast cancer cells (MDA-MB-231) in the cell incubator for 2 days, induce cell apoptosis, produce apoptotic body, collect the cell supernatant, record as cell Supernatant A: The incubator is sterile and non-toxic, the temperature is 35°C-37°C, and the medium is a complete medium containing 10% serum. A hypoxic environment can also be created by adding to the medium a compound that induces cellular hypoxia, such as cobalt chloride, defaminated ferric, and / or sodium sulfite, in an amount that is added to upregulate hypoxia-inducible factor 1α in tumor cells (HIF-1α).

[0079] 2. Extraction and purification of apoptotic bodies

[0080] (1) Removal of cells and cell debris: Centrifuge the cell supernatant A for 1 h under the condition of a centrifugal force of 50...

Embodiment 2

[0084] 1. Induce tumor cell apoptosis

[0085] In an oxygen-deprived environment (O 2 The concentration is 1%-6%), continue to culture breast cancer cells (MDA-MB-231) in the cell incubator for 2 days, induce cell apoptosis, produce apoptotic body, collect the cell supernatant, record as cell Supernatant A: The incubator is sterile and non-toxic, the temperature is 35°C-37°C, and the medium is a complete medium containing 10% serum.

[0086] 2. Extraction and purification of apoptotic bodies

[0087] (1) Removing cells and cell debris: Centrifuge the cell supernatant A for 30 min under the condition of a centrifugal force of 500 g, discard the precipitate, take the supernatant, and record it as supernatant B;

[0088] (2) Precipitate apoptotic bodies: Centrifuge the supernatant B for 1 h under the condition of a centrifugal force of 2500 g, discard the supernatant, and obtain the precipitate;

[0089] (3) Purification of apoptotic bodies: resuspend the precipitate obtained ...

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Abstract

The invention relates to a tumor cell apoptosis body and a preparation method and an application thereof. The preparation method of the tumor cell apoptosis body comprises the following steps: culturing tumor cells in an anoxic environment so as to induce the tumor cells to generate apoptosis and generate the apoptosis body. The tumor cell apoptosis body obtained by the preparation method has highactivity and excellent anti-tumor effect, can be used as a tumor vaccine for treating cancer, and has the advantages of accurate treatment targeting, low toxic and side effects and good treatment effect.

Description

technical field [0001] The invention relates to the field of anti-tumor vaccines, in particular to a tumor cell apoptosis body and its preparation method and application. Background technique [0002] In recent years, the incidence of malignant tumors has increased year by year. Although scientists have been exploring various ways to overcome tumors, the clinical cure rate of tumors has not yet shown a significant increase. Cancer treatment methods mainly include surgery, radiotherapy and drug therapy. However, these traditional cancer treatment methods are poorly targeted and highly toxic. While killing tumor cells, they will also damage normal cells and cause adverse reactions. Therefore, it has certain limitations. With the rapid development and cross penetration of related disciplines such as oncology, immunology, and molecular biology, the research on tumor immunotherapy has advanced by leaps and bounds. The US FDA has recently approved several immunotherapy projects...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/09A61K39/00A61K35/13A61P35/00
CPCA61K35/13A61K39/0011A61P35/00C12N5/0693C12N2500/02
Inventor 冯敏郭羚
Owner SUN YAT SEN UNIV
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