Exocrine body as well as preparation method and application thereof as tumor vaccine

A technology of exocrine and tumor cells, which is applied in a kind of exocrine and its preparation method and other fields, can solve the problems of unsatisfactory clinical curative effect, and achieve the effect of strong tumor immunogenicity, enhanced adhesion and high purity

Active Publication Date: 2015-12-09
BEIJING DOING TIMES BIOMEDICAL TECH
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] In the prior art, DC-tumor fusion cells are also directly used as tumor vaccines, but their clinical efficacy is often not very satisfactory.

Method used

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  • Exocrine body as well as preparation method and application thereof as tumor vaccine
  • Exocrine body as well as preparation method and application thereof as tumor vaccine
  • Exocrine body as well as preparation method and application thereof as tumor vaccine

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0045] Example 1 γδT-APC and NDV-Ulster virus strain modified tumor cells were fused, and the resulting fused cells For the preparation of exosomes

[0046] (1) Preparation of tumor cells modified by NDV-Ulster virus strain

[0047] The breast cancer cell line MCF7 was irradiated with gamma radiation to inactivate. The dose of gamma radiation was 200Gy. 7 Add NDV-Ulster virus strain to the ratio of 32 hemagglutination units of NDV-Ulster virus strain in each inactivated tumor cell to obtain a mixture, and the mixture is moved into a culture bottle, and a serum-free cell culture medium (manufacturer) is added in the culture bottle. Lonza, trade name X-VIVO15) and at 37 ° C, 5% CO 2 The tumor cells modified by NDV-Ulster were collected by centrifugation at 1500rpm×10min, washed with serum-free cell culture medium (manufacturer: Lonza, product name: X-VIVO15), and the cells were counted by trypan blue staining. The number and concentration should be tested for sterility. If...

Embodiment 2

[0057] Example 2 γδT-APC and NDV-Ulster virus strain modified tumor cells were fused, and the resulting fused cells for the preparation of exosomes

[0058] (1) Preparation of tumor cells modified by NDV-Ulster virus strain

[0059] The breast cancer cell line MCF7 was inactivated by adding mitomycin C so that the dose of mitomycin C was 50 μg / ml. 7 Add NDV-Ulster virus strain to the ratio of 32 hemagglutination units of NDV-Ulster virus strain in each inactivated tumor cell to obtain a mixture, and the mixture is moved into a culture bottle, and a serum-free cell culture medium (manufacturer) is added in the culture bottle. Takara, product name GT-T551H3) at 37 ° C, 5% CO 2 Cultivate under certain conditions for 2 hours, collect NDV-Ulster modified tumor cells by centrifugation at 1500rpm×10min, wash with serum-free cell culture medium (manufacturer is Takara, product name is GT-T551H3), and use trypan blue staining method to measure the cells The number and concentrati...

Embodiment 3

[0069] Example 3 Mo-DC cells were fused with unmodified tumor cells, and the resulting fused cells were used to prepare foreign secretory body

[0070] Induction culture of Mo-DC cells: purchase umbilical cord blood from the umbilical cord blood bank (the same umbilical cord blood as in Example 1), and separate the mononuclear cells with the human lymphocyte separation medium of GE Company with the trade name Ficoll-paquepremium, then use The serum-free cell medium of 10% described umbilical cord blood plasma (manufacturer is Lonza, trade name is X-VIVO15) mononuclear cell is diluted to 2 * 10 6 cells / ml, placed in a culture flask, 5% CO 2 , Culture at 37°C for 2 hours, gently suck out the suspended cells, retain the adherent cells in the lower layer, add GM-CSF with a final concentration of 100ng / ml, IL-4 with a final concentration of 100ng / ml, The umbilical cord blood plasma with a final concentration of 10%, the L-glutamine with a final concentration of 0.02mmol / L, and...

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Abstract

The invention discloses an exocrine body as well as preparation method and application thereof as a tumor vaccine, wherein the method comprises the following steps: (1) preparing NDV-Ulster virus strain modified tumor cells; (2) preparing novel phosphate antigen or biphosphonate drug stimulated and activated antigen presenting cells gamma-delta T-APC; (3) preparing a fusion cell of the gamma-delta T-APC and the NDV-Ulster modified tumor cells; (4) purifying the fusion cell; (5) culturing the fusion cell; (6) preparing a fusion cell exocrine body; and (7) preparing a vaccine. The invention has the following beneficial effects: more novel antigen presenting cells gamma-delta T-APC are prepared by the method, and a fusion rate between the gamma-delta T-APC and the modified tumor cells is higher; and the obtained exocrine body is more stable in antigen combination, more complete in tumor antigen titer and stronger in immunogenicity, and the exocrine body is high in fusion cell and exocrine body tumor antigen loading.

Description

technical field [0001] The present invention relates to an exocrine and its preparation method and the application of the exocrine as a tumor vaccine, in particular to a fusion preparation of tumor cells modified by novel antigen-presenting cells γδT-APC and Newcastle disease virus strain NDV-Ulser The method for preparing exosomes by using the fused cells, and also relates to the application of the obtained exosomes as tumor vaccines. Background technique [0002] Exosomes are vesicle structures that are released into the extracellular environment after the fusion of multivesicular body membranes and cell membranes in eukaryotic cells. Recent studies have found that various types of cells can release exosomes. Exosomes currently used to prepare tumor vaccines mainly come from the following sources: exosomes derived from tumor cells (hereinafter referred to as Texosomes) and dendritic cells (hereinafter referred to as Dendritic Cells, DCs) derived exosomes (hereinafter refe...

Claims

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Application Information

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IPC IPC(8): C12N5/22A61K39/00A61P35/00
Inventor 盖丽云李刚毅
Owner BEIJING DOING TIMES BIOMEDICAL TECH
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