Method for culturing natural killer cells using T cells
View Patent ↗The present invention relates to a method for producing natural killer cells using T cells, and more particularly, to a method for producing natural killer cells, which comprises culturing seed cells using CD4(+) T cells as feeder cells. The method for producing natural killer cells using T cells according to the present invention is a method capable of producing natural killer cells by selectively proliferating only natural killer cells from a small amount of seed cells while maintaining the high killing activity of the natural killer cells. The method of the present invention can produce a large amount of natural killer cells that can be frozen, and thus is useful for commercialization of cell therapeutic agents.
1. A method for treating cancer, the method comprising:
administering to a subject in need thereof a therapeutically effective amount of a population of natural killer cells produced by the method comprising:
(a) providing CD3(+)-depleted seed cells comprising mononuclear cells comprising natural killer cells;
(b) expanding the natural killer cells by culturing the seed cells with a plurality of cells from an inactivated CD4(+) T cell line in a medium comprising:
a T-cell stimulating antibody selected from the group consisting of OKT3, UCHT1, HTa, or a combination thereof; and
IL-2,
to produce expanded natural killer cells,
thereby producing the population of natural killer cells.
2. The method of claim 1 , wherein the seed cells provided in step (a) have not been expanded ex vivo or in vitro.
3. The method of claim 1 , wherein the seed cells provided in step (a) are selected from peripheral blood cells, peripheral blood leukocytes, and PBMCs (peripheral blood mononuclear cells).
4. The method of claim 1 , wherein the cells from an inactivated CD4(+) T cell line are selected from the group consisting of H9, HuT78, Molt-3, PEER, and combinations thereof.
5. The method of claim 4 , wherein the cells from an inactivated CD4(+) T cell line are selected from the group consisting of H9, HuT78, and combinations thereof.
6. The method of claim 1 , wherein the culturing is carried out for 5-60 days.
7. The method of claim 1 , further comprising:
(c) culturing the expanded natural killer cell(s) produced in step (b) with a second plurality of cells from an inactivated CD4(+) T cell line.
8. The method of claim 1 , wherein the population of natural killer cells is administered as part of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.
9. The method of claim 1 , wherein the cancer is selected from blood cancer, lymphoma, liver cancer, brain cancer, retinoblastoma, neuroblastoma, and ovarian cancer.
10. The method of claim 7 , wherein the second plurality of cells from an inactivated CD4(+) T cell line are selected from the group consisting of H9, HuT78, Molt-3, PEER, and combinations thereof.
11. The method of claim 7 , wherein the second plurality of cells from an inactivated CD4(+) T cell line are selected from the group consisting of H9, HuT78, and combinations thereof.
12. The method of claim 5 , wherein the seed cells provided in step (a) have not been expanded ex vivo or in vitro.
13. The method of claim 12 , wherein the population of natural killer cells is administered as part of a pharmaceutical composition further comprising a pharmaceutically acceptable carrier.
14. The method of claim 13 , wherein the cancer is selected from blood cancer, lymphoma, liver cancer, brain cancer, retinoblastoma, neuroblastoma, and ovarian cancer.