A tumor drug-loaded microparticle preparation and its preparation method
A micro-particle and drug-carrying technology, applied in anti-tumor drugs, pharmaceutical formulations, drug combinations, etc., to achieve the effect of improving killing effect, simple operation and low cost
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Embodiment 1
[0051] Example 1: Tumor drug-loaded microparticle preparations produced by using chemotherapeutic drugs to induce apoptosis of mouse liver cancer stem cells
[0052] 1. Experimental materials and reagents
[0053] H22 mouse liver cancer cells, doxorubicin (with red fluorescence), carboxyfluorescein succinimidyl ester (CFSE) (green fluorescent dye).
[0054] 2. Experimental steps
[0055] 1) Cultivate H22 mouse liver cancer cells in RPMI 1640 cell culture medium, and the H22 cells are pre-diluted to 1×10 with RPMI 1640 medium 4 cells / mL to obtain a cell suspension, dilute the soft fibrin glue to a 2mg / mL solution with T7 buffer, and mix the solution with the H22 cell suspension at a volume ratio of 1:1 to obtain a mixture; Add 1 μL of thrombin (0.1U / μL) to a 96-well plate, and mix it with 50 μL of the above mixture to obtain a mixed solution. After incubating at 37°C for 15 minutes, add the mixed solution to 200 μL of RPMI 1640 for complete culture The tumor stem cell clones...
Embodiment 2
[0061] Example 2: The drug-loaded microparticles were obtained after incubating the microparticles produced by the apoptosis of mouse liver cancer stem cells induced by ultraviolet rays and chemotherapeutic drugs.
[0062] 1. Experimental materials and reagents
[0063] The H22 mouse liver cancer cells used were the same as in Example 1, the ultraviolet device was owned by a conventional cell clean bench, and the doxorubicin was the same as in Example 1.
[0064] 2. Experimental steps
[0065] 1) Culture H22 mouse liver cancer stem cells to make the cell volume reach 2×10 7 , culture method with embodiment 1.
[0066] 2) The H22 mouse liver cancer stem cells (with cell culture medium) were irradiated with ultraviolet light for 60 min.
[0067] 3) 24 hours after ultraviolet irradiation, the supernatant of the apoptotic tumor stem cell culture solution was gradually centrifuged, and the method was the same as in Example 1 to obtain microparticles of the tumor stem cell.
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Embodiment 3
[0071] Example 3: Accumulation of drug-loaded microparticles in tumor tissue
[0072] 1. Experimental materials and reagents
[0073] The H22 mouse liver cancer cells used were the same as in Example 1, the chemotherapeutic drug doxorubicin was commercially obtained, and the BALB / C mice were purchased from the Medical Animal Center of Wuhan University.
[0074] 2. Experimental steps
[0075] 1) Culture H22 mouse liver cancer cells and liver cancer stem cells, so that the cell volume reaches 2×10 7 , wherein the culture method of H22 mouse liver cancer stem cells is the same as in Example 1, and the H22 mouse liver cancer cells are cultured with RPMI 1640 medium.
[0076] 2) Take 1×10 from the above cultured cells 7 Doxorubicin was administered to one cell to make the final concentration of doxorubicin reach 200 μg / mL, and the remaining cells continued to be cultured without any treatment.
[0077] 3) 24 hours after the administration of chemotherapeutic drugs, the drug-loa...
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