Method for induction of T cells from pluripotent stem cells
The present invention provides a method for efficiently inducing CD8-positive T cells by adding vitamin C to the medium in the steps of induction of the CD8-positive T cells from pluripotent stem cells. The present invention also provides a method for efficiently inducing CD8-positive T cells by performing culture in a medium supplemented with an adrenocortical hormone agent in the step of induction of the CD8-positive T cells from CD4/CD8 double-positive T cells.
1. A method for inducing CD8-positive T cells, comprising the step of culturing CD4/CD8 double-positive T cells in a medium supplemented with an adrenocortical hormone agent and IL-7, wherein the adrenocortical hormone agent is glucocorticoid or a derivative thereof.
2. The method according to claim 1 , wherein prior to said step of culturing the CD4/CD8 double-positive T cells in a medium supplemented with said adrenocortical hormone agent and interleukin (IL)-7, the method comprises the steps of:
(1) culturing pluripotent stem cells in a medium supplemented with vitamin C and vascular endothelial growth factor (VEGF) to induce hematopoietic progenitor cells; and
(2) culturing cells obtained in said Step (1) in a medium supplemented with vitamin C, FLT-3L and IL-7 to induce the CD4/CD8 double-positive T cells.
3. The method according to claim 2 , wherein said vitamin C is vitamin C phosphate.
4. The method according to claim 2 , wherein the vitamin C is supplied to the medium every day.
5. The method according to claim 2 , wherein, in said Step (1), the pluripotent stem cells are cultured on C3H10T1/2 cells.
6. The method according to claim 2 , wherein said Step (1) is carried out under low oxygen conditions with an oxygen concentration of not more than 5%.
7. The method according to claim 2 , wherein, in said Step (1), the medium further comprises Stem cell factor (SCF), and Flt3 Ligand (FLT-3L).
8. The method according to claim 2 , wherein, in said Step (2), the cells obtained in Step (1) are cultured on OP9-DL1 cells.
9. The method according to claim 1 , wherein said adrenocortical hormone agent is dexamethasone.
10. The method according to claim 1 , wherein said medium further comprises an anti-CD3 antibody, vitamin C, and IL-2.
11. The method according to claim 10 , wherein said vitamin C is vitamin C phosphate.
12. The method according to claim 2 , wherein, in said Step (1), the pluripotent stem cells are cultured on feeder cells.
13. The method according to claim 2 , wherein, in said Step (2), the cells obtained in Step (1) are cultured on feeder cells.
14. The method according to claim 1 , wherein said medium further contains a T cell stimulant.
15. The method according to claim 1 , wherein the adrenocortical hormone agent is selected from the group consisting of cortisone acetate, hydrocortisone, fludrocortisone acetate, prednisolone, triamcinolone, methylprednisolone, dexamethasone, betamethasone, and beclometasone dipropionate.