Method for producing CD4/CD8 double-positive T cells
A method for producing CD4/CD8 double-positive T cells, comprising the steps of: (1) culturing pluripotent stem cells in a medium to induce hematopoietic progenitor cells; and (2) culturing the hematopoietic progenitor cells obtained in the step (1) in a medium containing a p38 inhibitor and/or SDF-1 to induce CD4/CD8 double-positive T cells.
1. A method for producing CD4/CD8 double-positive T cells, the method comprising the step of culturing hematopoietic progenitor cells in a medium containing a p38 inhibitor and SDF-1 and further containing a delta ligand, SCF, TPO, FLT-3L, and IL-7 to induce CD4/CD8 double-positive T cells, wherein the step is carried out by adherent culture.
2. The method according to claim 1 , wherein the p38 inhibitor is SB203580.
3. The method according to claim 1 , wherein, in the step, the medium contains vitamin C.
4. The method according to claim 1 , wherein the hematopoietic progenitor cells are cultured without using feeder cells.
5. The method according to claim 1 , wherein the hematopoietic progenitor cells are hematopoietic progenitor cells differentiated from pluripotent stem cells.
6. The method according to claim 5 , wherein the pluripotent stem cells are induced pluripotent stem cells.
7. The method according to claim 6 , wherein the induced pluripotent stem cells are derived from somatic cells other than T cells.
8. The method according to claim 6 , wherein the induced pluripotent stem cells are induced pluripotent stem cells having a chimeric antigen receptor introduced therein.
9. A method for producing CD4/CD8 double-positive T cells wherein the step is carried out by adherent culture, comprising the steps of:
(1) culturing pluripotent stem cells in a medium to induce hematopoietic progenitor cells; and
(2) culturing the hematopoietic progenitor cells obtained in the step (1) in a medium containing a p38 inhibitor and SDF-1 and further containing a delta ligand, SCF, TPO, FLT-3L, and IL-7 to induce CD4/CD8 double-positive T cells.
10. The method according to claim 9 , wherein the p38 inhibitor is SB203580.
11. The method according to claim 9 , wherein the step (1) is carried out by suspension culture.
12. The method according to claim 9 , wherein, in the steps (1) and (2), the medium contains vitamin C.
13. The method according to claim 9 , wherein, in the steps (1) and (2), the culture is carried out without using feeder cells.
14. The method according to claim 9 , wherein the pluripotent stem cells are induced pluripotent stem cells.
15. The method according to claim 14 , wherein the induced pluripotent stem cells are derived from somatic cells other than T cells.
16. The method according to claim 14 , wherein the induced pluripotent stem cells are induced pluripotent stem cells comprising a transgene encoding a chimeric antigen receptor.