Tumor vaccine and preparation method thereof
A tumor vaccine and tumor cell technology, applied in vaccines, anti-tumor drugs, pharmaceutical formulations, etc., can solve the problems of ineffectiveness of tumor antigens, poor targeting and use safety, etc., achieve a broad spectrum of tumor antigens, and enhance targeted lethality , the effect of improving efficiency
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Embodiment 1
[0049] Example 1: Tumor cells produce cell vesicles after undergoing apoptosis treatment.
[0050] 1. Experimental materials and reagents
[0051] H22 mouse liver cancer cells, fluorescent dye: carboxyfluorescein diacetate succinimide ester (commercially available, with green fluorescence), and the ultraviolet device is owned by a conventional cell ultra-clean bench.
[0052] 2. Experimental steps
[0053] 1) Culture H22 mouse liver cancer cells in DMEM cell culture medium to make the cell volume reach 2×10 7 ;
[0054] 2) After the H22 mouse liver cancer cells were stained with carboxyfluorescein diacetate succinimide lipid fluorescent dye, fresh culture medium was added, and then the above H22 mouse liver cancer cells were divided into two groups (H22-1 group, H22-1 group, H22-2 group), wherein one group of H22-1 was irradiated with ultraviolet light for 60 minutes, and the other group of H22-2 was not treated and cultured normally in the culture medium;
[0055] 3) 48 h...
Embodiment 2
[0060] Example 2: Cell vesicles produced by induced tumor cells can be taken up by dendritic cells.
[0061] 1. Experimental materials and reagents
[0062] The H22 mouse liver cancer cells and the ultraviolet device used were the same as in Example 1, carboxyfluorescein diacetate succinimidyl ester (CFSE) (green fluorescent dye), PKH26 (red fluorescent dye).
[0063] 2. Experimental steps
[0064] 1) Cultivate H22 mouse liver cancer cells according to the method described in Example 1, so that the cell volume reaches 1×10 7 ; Use carboxyfluorescein diacetate succinimide lipid to stain the above H22 mouse liver cancer cells, then add fresh culture medium, and use ultraviolet radiation for 60 minutes. After 48 hours of ultraviolet radiation, H22 mouse liver cancer cells appear Obviously smaller and dim state, confirm that these tumor cells have been induced apoptosis by ultraviolet rays, and collect the cell vesicles induced by ultraviolet rays with green fluorescence accordi...
Embodiment 3
[0069] Example 3: Dendritic cells up-regulate the expression of co-stimulatory signal molecules after taking up cell vesicles.
[0070] 1. Experimental materials and reagents
[0071] The H22 mouse hepatoma cells used were the same as in Example 1, the dendritic cells were the same as in Example 2, and the fluorescently labeled antibodies against mouse CD80, CD86, and MHC II.
[0072] 2. Experimental steps
[0073] 1) The culture method of dendritic cells is the same as that in Example 2; the cell vesicles of H22 mouse liver cancer cells are prepared by the method in Example 1, and the method for making dendritic cells ingest H22 mouse liver cancer cells is the same as in Example 2.
[0074] 2) The cell vesicles and dendritic cells of H22 mouse liver cancer cells were incubated for 48 hours, and the dendritic cells that had taken up the cell vesicles were used as the experimental group; at the same time, the individual dendritic cells were incubated for 48 hours, as a contro...
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