Method for producing dendritic cells
View Patent ↗Disclosed are embryonic stem cell-derived dendritic cells, genetically modified immature dendritic cells capable of maturation, as well as methods for the production of such cells. In one embodiment, the cells made be produced by a method comprising the steps of providing a population of embryonic stem cells; culturing the embryonic stem cells in the presence of a cytokine or combination of cytokines which brings about differentiation of the embryonic stem cells into dendritic cells; and recovering the dendritic cells from the culture. In a further embodiment, the cells may be genetically modified.
1. A process for producing a culture of human dendritic cells, comprising: culturing embryoid bodies made from human embryonic stem cells, said culturing being in the presence of a composition comprising IL-3; and recovering human dendritic cells from said culture.
2. The process according to claim 1 , wherein said composition further comprises GM-CSF.
3. The process according to claim 1 , wherein said human embryonic stem cells are genetically modified.
4. The process of claim 3 , wherein said human embryonic stem cells are transfected with a gene, which gene is expressed in the human dendritic cells.
5. The process of claim 4 , wherein the gene is under the control of a promoter which is preferentially active in dendritic cells.
6. The process of claim 4 , wherein the gene is a reporter gene which expresses a detectable protein in the human dendritic cells.
7. The process of claim 6 , wherein the detectable protein is a fluorescent protein.
8. The process of claim 7 , wherein the fluorescent protein is a green fluorescent protein (GFP).
9. The process of claim 3 , wherein the human embryonic stem cells are genetically modified so as to inactivate a copy of a gene.
10. The process of claim 1 , wherein the human dendritic cells express one or more heterologous gene(s).
11. The process of claim 10 , wherein the heterologous gene(s) encodes a cell surface protein.
12. The process of claim 10 , wherein the heterologous gene(s) encodes Fas-ligand.
13. The process of claim 10 , wherein the heterologous gene(s) encodes an antigen.
14. The process of claim 13 , wherein the antigen is an autoantigen, a tumour antigen, or a foreign antigen.
15. The process of claim 13 , wherein the antigen is a tumour antigen.
16. The process of claim 10 , wherein the heterologous gene(s) is an anti-apoptotic gene.
17. The process of claim 10 , wherein the heterologous gene(s) encodes FLIP or bcl-2.
18. The process of claim 1 , wherein the human dendritic cells co-express two or more heterologous genes.
19. The process of claim 1 , wherein one or more endogenous gene(s) has been inactivated in the human embryonic stem cells.
20. The process of claim 19 , wherein the inactivated endogenous gene(s) is B7-1, IL-12, p35 subunit of IL-12 or p40 subunit of IL-12.
21. The process of claim 1 , wherein the recovered human dendritic cells are substantially pure.
22. A process for producing a culture of mouse dendritic cells, comprising: culturing embryoid bodies made from mouse embryonic stem cells, said culturing being in the presence of a composition comprising IL-3; and recovering mouse dendritic cells from said culture.
23. The process according to claim 22 , wherein the composition further comprises GM-CSF.
24. The process according to claim 22 , wherein said mouse embryonic stem cells are genetically modified.
25. The process of claim 24 , wherein said mouse embryonic stem cells are transfected with a gene, which gene is expressed in the mouse dendritic cells.
26. The process of claim 25 , wherein the gene is under the control of a promoter which is preferentially active in dendritic cells.
27. The process of claim 25 , wherein the gene is a reporter gene which expresses a detectable gene product in the mouse dendritic cells.
28. The process of claim 27 , wherein the detectable gene product is a fluorescent protein.
29. The process of claim 28 , wherein the fluorescent protein is a green fluorescent protein (GFP).
30. The process of claim 24 , wherein the mouse embryonic stem cells are genetically modified so as to inactivate a copy of a gene.
31. The process of claim 22 , wherein the mouse dendritic cells express one or more heterologous gene(s).
32. The process of claim 31 , wherein the heterologous gene(s) encodes a cell surface protein.
33. The process of claim 31 , wherein the heterologous gene(s) encodes an antigen.
34. The process of claim 22 , wherein the mouse dendritic cells co-express two or more heterologous genes.
35. The process of claim 22 , wherein one or more endogenous gene(s) has been inactivated in the mouse embryonic stem cells.
36. The process of claim 22 , wherein the recovered dendritic cells are substantially pure.
37. The process of claim 22 , wherein the mouse embryonic stem cells are from a CBA/Ca or a C57Bl/6 cell line.
38. The process of claim 22 , wherein the mouse embryonic stem cells are from the ESF116 cell line.