Colorectal cancer stem cell vaccine and its preparation method and application
A colorectal cancer, stem cell technology, applied in the direction of tumor/cancer cells, animal cells, vertebrate cells, etc., can solve the problems of low efficacy and lack of colorectal cancer stem cell vaccine, etc., to enhance the efficacy, improve the body's immune function, and easily The effect of quality control
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Embodiment 1
[0041] Example 1: Preparation of colorectal cancer stem cell vaccine
[0042] In this example, colorectal cancer stem cell vaccines were prepared using colorectal cancer cells stored in liquid nitrogen as raw materials.
[0043] 1. Put the colorectal cancer cell HCT-15 stored in liquid nitrogen or -80°C in a 37°C water bath to dissolve quickly;
[0044] 2. Mix the thawed tumor cell suspension with 8.0ml medium (RPMI1640+10% FBS), and centrifuge at 1500 rpm for 8 minutes;
[0045] 3. Discard the supernatant, then absorb 8.0ml culture medium to mix the cell pellet, then centrifuge at 1500 rpm for 8 minutes, discard the supernatant, mix the cell pellet with 1.0ml culture medium, and set aside;
[0046] 4. Take another 75cm 2 Square bottle, add 14.0ml of culture medium, add the suspension containing tumor cells (1.0ml) prepared above into this square bottle, at 37°C, 5% CO 2 cultured in an incubator;
[0047] 5. Tumor cells grow on the wall. After 3-4 days, when the tumor cell...
Embodiment 2
[0064] Example 2: Observation of curative effect after injection of colorectal cancer stem cell vaccine
[0065] The colorectal cancer stem cell vaccine prepared in Example 1 was applied to the treatment of colorectal cancer patients. The detailed experimental process was as follows: 30 colorectal cancer patients were randomly selected and divided into two groups, 15 of which received the vaccine prepared in Example 1. The vaccine injection treatment was used as the treatment group; the other 15 people who did not receive the vaccine treatment were used as the control group. Before injecting patients, 5ml of blood was drawn from each patient, and the number of peripheral blood lymphocytes (T BNK detection kit) and lymphocyte function (6colour T BNK With Trucount detection kit) were detected, and the report sheet was obtained through flow cytometry detection. The patients in the treatment group were subcutaneously injected with a vaccine dose of 500ul into the colorectal cancer...
Embodiment 3
[0070] Example 3: Humoral immune response induced by the tumor stem cell vaccine prepared in Example 1
[0071] 30 routine patients in the embodiment 1 are equally divided into two groups (no vaccine injection group-control group and vaccination group-experimental group), carry out according to the following method respectively:
[0072] Collect 5ml of the whole blood of the patient on the 14th day, and separate the plasma with the lymphocyte separation medium. The details are as follows: Take 5ml of fresh anticoagulant blood, mix it with normal saline for injection at a ratio of 1:1, and carefully add it to 2ml of the lymphocyte separation medium , centrifuge at 400g (horizontal rotor with a radius of 15cm) for 20 minutes. At this time, the centrifuge tube is divided into four layers, and the uppermost layer is plasma. The IgG content in plasma was detected by ELISA.
[0073] The result is as image 3 As shown, compared with the control group, P image 3 It can be seen that ...
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