Method for the induction and expansion of natural killer cells derived from peripheral blood mononuclear cells
The present invention relates to a method for inducing and expanding natural killer cells derived from peripheral blood mononuclear cells, which comprises co-culturing, as feeder cells, irradiated Jurkat cells and irradiated Epstein-Barr virus transformed lymphocyte continuous line (EBV-LCL) cells in the presence of cytokines, along with peripheral blood mononuclear cells. According to the present invention, a large quantity of natural killer cells can be induced and proliferated from a small quantity of peripheral blood mononuclear cells even without the use of high-cost equipment or various kinds of expensive cytokines, thereby making it possible to significantly improve the efficiency and efficacy of the prevention and treatment of cancer using the natural killer cells.
1 . A method for treating a cancer in a subject in need thereof, wherein the method comprises:
expanding natural killer cells in a co-culture comprising peripheral blood mononuclear cells and feeder cells, wherein the feeder cells comprise irradiated Jurkat cells and irradiated Epstein-Barr virus transformed lymphocyte continuous cell line (EBV-LCL) cells and the co-culture is in the presence of one or more cytokines; and
administering an effective amount of a composition comprising the expanded natural killer cells to the subject, thereby treating the cancer.
2 . The method of claim 1 , wherein the one or more cytokines is selected from IL-2, IL-15, IL-21, Flt3-L, SCF, or IL-7.
3 . The method of claim 2 , wherein the one or more cytokines is IL-2.
4 . The method of claim 3 , wherein the IL-2 is present at a concentration of 500 U/ml.
5 . The method of claim 1 , wherein the peripheral blood mononuclear cells are obtained from a normal person or a person suffering from the cancer.
6 . The method of claim 1 , wherein the peripheral blood mononuclear cells are obtained from the subject.
7 . The method of claim 1 , wherein the peripheral blood mononuclear cells are obtained from a healthy subject.
8 . The method of claim 1 , wherein each of the irradiated Jurkat cells or the irradiated EBV-LCL cells is obtained by treatment with radiation of 100 to 500 Gy.
9 . The method of claim 1 , wherein the peripheral blood mononuclear cells are co-cultured for 5 days to 14 days.
10 . The method of claim 1 , wherein the peripheral blood mononuclear cells and feeder cells are mixed at a ratio of 1:0.5:0.5 during the co-culturing.
11 . The method of claim 1 , wherein the co-culturing comprises adding the irradiated Jurkat cells, the irradiated EBV-LCL cells and the one or more cytokines on the 1st to 15th day.
12 . The method of claim 1 , wherein the cancer comprises lung cancer, liver cancer, breast cancer, or blood cancer.
13 . A method for treating a cancer in a subject in need thereof, the method comprising:
providing a composition comprising natural killer cells, wherein providing the composition comprises expanding natural killer cells in a co-culture comprising peripheral blood mononuclear cells and feeder cells for 5 days to 14 days,
wherein the feeder cells comprise irradiated Jurkat cells and irradiated Epstein-Barr virus transformed lymphocyte continuous cell line (EBV-LCL) cells,
wherein the co-culture is in the presence of one or more cytokines, wherein the one or more cytokines comprise IL-2, and
wherein the peripheral blood mononuclear cells and feeder cells are mixed at a ratio of 1:0.5:0.5 during the co-culturing; and
administering an effective amount of the composition to the subject, thereby treating the cancer.
14 . The method of claim 13 , wherein the one or more cytokines are IL-2.
15 . The method of claim 13 , wherein the cancer comprises lung cancer, liver cancer, breast cancer, or blood cancer.