Perfusion culture method for car-t cells

Abstract

Provided is a perfusion culture method for CAR-T cells. The method comprises the following steps: 1) separating peripheral blood mononuclear cells from a single blood cell of a subject, and sorting the mononuclear cells to obtain T cells; 2) carrying out activation treatment on the separated T cells by using CD3/CD28 stimulation magnetic beads; 3) infecting the activated T cells by using a lentiviral vector; 4) carrying out perfusion culture on the lentivirus-infected T cells, and harvesting CAR-T cells, wherein the composition of a serum-free culture medium, which does not contain an animal-derived component, for culturing the CAR-T cells is: AIM-V+(3-9)% ISR; and the perfusion culture comprises the following stages: the first stage: when the cell density is (0.5-1.1)×106 cells/mL, the perfusion rate is A1, and/or the second stage: when the cell density is (1.1-2)×106 cells/mL, the perfusion rate is A2, and the third stage: when the cell density is >2×106 cells/mL, the perfusion rate is A3, and the ratio of A1 to A2 to A3 is 1:2:2.5.

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