Preparation method of composite immune cell with high killing capability on tumor cells
A technology of immune cells and tumor cells, applied in the field of compound immune cell preparation, which can solve the problems of poor infiltration of solid tumors, inability to penetrate deep into tumors, and short retention in the body
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[0042] like Figure 5 As shown, a method for preparing composite immune cells with high killing ability to tumor cells includes the following steps in sequence:
[0043] Step 1, collect peripheral blood and isolate mononuclear cells, culture the isolated mononuclear cells, and obtain suspension cells and adherent cells;
[0044] Step 2, culturing mature DC cells through the adherent cells;
[0045] Step 3, inoculating the suspension cells with a specific immune cell basal culture medium to cultivate a specific immune cell culture system;
[0046] Step 4, adding the DC cells obtained in the step 2 to the specific immune cell culture system for co-cultivation.
[0047] According to a preferred embodiment of the present invention, the specific immune cells are selected from at least one of γδ T cells and NKT / NK cells.
[0048] According to a preferred embodiment of the present invention, the step 3 further includes the following steps:
[0049] Step 301, adding corresponding ...
Embodiment 1
[0063] Example 1 presents a preparation method and corresponding experimental effects of a composite immune cell for the treatment of cancer.
[0064] It includes the following steps:
[0065] 1. Isolation of mononuclear cells:
[0066] 1.1 Aseptic intravenous collection of 100ml of peripheral blood from healthy adult volunteers for cell preparation, the collection of blood samples conforms to the principle of informed consent;
[0067] 1.2 Add 15ml of Lymphoprep human lymphocyte separation solution dropwise to the centrifuge tube, and slowly add whole blood to the upper layer of the separation solution along the tube wall to avoid blood infiltration to the lower layer of the separation solution;
[0068] 1.3 Centrifuge at 900 × g for 20 min at room temperature, adsorb the upper plasma into another centrifuge tube, inactivate at 56 °C, let it stand in the refrigerator at minus 20 °C for 10 min, and then use the supernatant after centrifugation again;
[0069] 1.4 Aspirate th...
Embodiment 2
[0095] Example 2 shows the effect of DC-γδT-NK / NKT cell mixture on three cancer cell lines.
[0096] Hela, K562, and SK-MES-1 cells were stained with 5 μM CFSE, DC-NK / NKT and γδT cell mixture or PBS and cancer cells were mixed in different proportions, and after 4 h incubation at 37 °C, 1 μg / ml of PI dye, CFSE+PI double positive cells are dead cells, such as image 3 shown, image 3 The data comes from the flow cytometry killing test. The high-efficiency target ratio DC-γδT-NK / NKT cell mixture can significantly enhance the killing of cancer cells. In terms of the effector-target ratio of the immune cell mixture and cancer cells in the first group, a higher killing rate was achieved, especially in the 4:1 effector-target ratio group, the killing rate of Hela, K562, and SK-MES-1 reached 74.6%, respectively. 70.2%, 68.1%. Shows a good killing effect on various types of cancer cells.
[0097] It can be concluded from the above examples that through the above co-cultivation mod...
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