Method for amplifying NK cells
A technique is needed which can amplify NK cells in vitro and prepare optimum number of NK cells for the adoptive immunotherapy. A method for amplifying NK cells is provided which comprises steps of: preparing cell population which is comprised of NK cells, removing T cells from the cell population which is comprised of NK cells, and, after removal of T cells, cultivating the remaining cells in a medium supplemented with 2500 to 2831 IU/mL of IL-2. The method for amplifying NK cells of the present invention may comprise a step of removing hematopoietic progenitor cells from the cell population. The present invention provides a pharmaceutical composition for adoptive immunotherapy, comprising NK cells which are prepared by the amplifying method of the present invention.
1. A method for amplifying NK cells, comprising the steps of:
preparing cell population which is comprised of NK cells;
removing T cells from the cell population which is comprised of NK cells; and
after removal of T cells, cultivating the remaining cells without feeder cells in a medium supplemented with 2500 to 2831 IU/mL of IL-2, as the only cytokine.
2. The method according to claim 1 , wherein the step of removing T cells is implemented by a step of removing CD3-positive cells.
3. The method according to claim 1 , comprising, between the step of preparing and the step of cultivating a step of:
removing hematopoietic progenitor cells from the cell population.
4. The method according to claim 3 , wherein the step of removing hematopoietic progenitor cells from the cell population is implemented by a step of removing CD34-positive cells.
5. The method according to claim 1 , wherein the medium comprise self serum of the donor, AB-type serum, and/or serum albumin.
6. The method according to claim 1 , wherein the step of preparing cell population which is comprised of NK cells is implemented by a step of separating mononuclear cells from blood cells collected from a subject.
7. The method according to claim 6 , wherein the blood cells are collected from peripheral blood, umbilical cord blood, a bone marrow and/or a lymph node.
8. The method according to claim 7 , wherein the blood cells are collected from peripheral blood using apheresis.
9. The method according to claim 1 , wherein the cell population is prepared from at least one kind of cells selected from a group consisting of:
hematopoietic stem cells derived from any stem cells selected from a group consisting: embryonic stem cells, adult stem cells and induced pluripotent stem cells (iPS cells);
hematopoietic stem cells derived from umbilical cord blood;
hematopoietic stem cells derived from peripheral blood;
hematopoietic stem cells derived from bone marrow blood;
umbilical cord blood mononuclear cells; and
peripheral blood mononuclear cells.
10. A method for adoptive immunotherapy comprising the steps of:
preparing a cell population which is comprised of NK cells,
removing T cells from the cell population,
after removal of T cells, cultivating the remaining cells without feeder cells in a medium supplemented with 2500 to 2831 IU/mL of IL-2, as the only cytokine, and
transplanting the NK cells which are amplified from the remaining cells to a patient.
11. The method according to claim 10 , wherein the step of removing T cells is implemented by a step of removing CD3-positive cells.
12. The method according to claim 10 , comprising, between the step of preparing and the step of cultivating, a step of:
removing hematopoietic progenitor cells from the cell population.
13. The method according to claim 12 , wherein the step of removing hematopoietic progenitor cells from the cell population is implemented by a step of removing CD34-positive cells.
14. The method according to claim 10 , wherein the medium comprise self serum of the donor, AB-type serum, and/or serum albumin.
15. The method according to claim 10 , wherein the step of preparing cell population which is comprised of NK cells is implemented by a step of separating mononuclear cells from blood cells collected from a subject.
16. The method according to claim 15 , wherein the blood cells are collected from peripheral blood, umbilical cord blood, a bone marrow and/or a lymph node.
17. The method according to claim 16 , wherein the blood cells are collected from peripheral blood using apheresis.
18. The method according to claim 10 , wherein the cell population is prepared from at least one kind of cells selected from a group consisting of:
hematopoietic stem cells derived from any stem cells selected from a group consisting: embryonic stem cells, adult stem cells and induced pluripotent stem cells (iPS cells);
hematopoietic stem cells derived from umbilical cord blood;
hematopoietic stem cells derived from peripheral blood;
hematopoietic stem cells derived from bone marrow blood;
umbilical cord blood mononuclear cells; and
peripheral blood mononuclear cells.