The invention relates to a CAR-NK
cell, and a preparation method and an application thereof. The preparation method comprises the following steps: S1, synthesizing a CAR sequence; S2, integrating theCAR sequence into a lentiviral target vector
plasmid to obtain a target
plasmid; S3, mixing the target
plasmid with a 293
T cell, performing culturing, collecting the obtained
virus liquid, and concentrating the
virus liquid to obtain the
virus concentrate of the target plasmid; S4, collecting
autologous plasma and
peripheral blood mononuclear cells (PBMC), adjusting the
cell density with an activation culture medium, performing
cell culturing, transferring obtained cells into a new cell culturing bag, continuing culturing, adding a proliferation culturing medium, and obtaining the NK cell after the culturing is finished; and S5, adding the virus concentrate of the target plasmid to the NK cell, and performing incubation to obtain the CAR-NK cell. The CAR-NK cell has very strong factor
secretion ability and
tumor inhibition ability, and the
transfection method for preparing the CAR-NK cell is efficient and stable.