Preparation of high efficiency immune active cell and method of using for anti tumour
A technology of immune activity and cell population, applied in the biological field, can solve the problems of hemorrhage, need large dose, difficult to obtain tumor antigen, etc., and achieve the effect of high proliferative activity and high cytotoxic activity
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[0032] The present invention will be further described below in conjunction with specific embodiments.
[0033] 1. Preparation of peripheral blood mononuclear cells:
[0034] Take 35ml of anticoagulated peripheral blood of tumor patient, dilute it in two-fold with normal saline, and separate PBMC with lymphocyte separation liquid with specific gravity of 1.077. The centrifuge rotor used for separation is a horizontal rotor with a centrifugal speed of 2000rpm and centrifugation for 20 minutes. Then wash the PBMC twice with Hanks balance solution, and then count the cells under the microscope, and finally adjust the cell density to 4×10-6 / ml cell suspension with lymphocyte culture medium completely F12 / DMEM 1:1 or AIM-V.
[0035] 2. Separation of peripheral blood lymphocytes and adherent cells:
[0036] Transfer the cell suspension into a 6-well plate, add 4ml to each well, set 37C, 5% CO 2 Incubate in a saturated humidity incubator for 2.5 hours, then gently wash the cells with a p...
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