Ex vivo generation of functional leukemia cells in a three-dimensional bioreactor
The present invention provides cultured leukemia cells. The method comprises isolating mononuclear cells, which contain leukemia cells, culturing the leukemia cells in a chamber having a scaffolding covered or surrounded with culture medium, where the scaffolding allows for leukemia cells to have cell to cell contacts in three dimensions. The subject leukemia cells are useful for screening compounds which inhibit or stimulate leukemia cell function or formation.
1. A method of culturing leukemia cells, comprising:
isolating mononuclear cells, which contain leukemia cells, from a subject and
culturing the cells in a chamber having a scaffolding covered or surrounded with culture medium, wherein said scaffolding allows for leukemia cells to have cell to cell contacts in three dimensions, and the leukemia cells are cultured for a duration of at least four weeks.
2. The method according to claim 1 , wherein the leukemia cells are mammalian leukemia cells.
3. The method according to claim 2 , wherein the mammalian leukemia cells are human leukemia cells.
4. The method according to claim 1 , wherein the leukemia cells are acute myelocytic cells.
5. The method according to claim 1 , wherein the leukemia cells are chronic lymphocytic leukemia cells.
6. The method according to claim 1 , wherein the scaffolding is selected from the group consisting of tangled fibers, porous particles, sponge, sponge-like material, and combinations thereof.
7. The method according to claim 1 , wherein the scaffolding is formed from a material selected from the group consisting of metal, glass, ceramic, plastic, hydroxyapatite, treated or untreated bone, a synthetic polymer, a natural substance, a semisynthetic material, and combinations thereof.
8. The method according to claim 7 , wherein the material is degradable.
9. The method according to claim 7 , wherein the material is non-degradable.
10. The method according to claim 1 , wherein the culture medium comprises exogenous growth factors.
11. The method according to claim 1 , wherein the culture medium includes granulocyte colony stimulating factor, granulocyte monocyte colony stimulating factor, IL3, or IL2.
12. The method according to claim 1 further comprising:
reseeding the chamber with mononuclear cells.
13. The method according to claim 1 , wherein the mononuclear cells are from peripheral blood, bone marrow aspirate or biopsy, chloroma, or spleen.
14. A method of culturing leukemia cells, comprising:
isolating mononuclear cells, which contain leukemia cells, from a subject and
culturing the cells in a chamber having a scaffolding covered or surrounded with culture medium, wherein said scaffolding is selected from the group of materials consisting of tangled fibers, porous particles, sponge, sponge-like material, and combinations thereof, wherein the leukemia cells must be able to enter into openings of the fibrous or porous materials, and the scaffolding allows for leukemia cells to have cell to cell contacts in three dimensions.
15. The method according to claim 14 , wherein the leukemia cells are mammalian leukemia cells.
16. The method according to claim 15 , wherein the mammalian leukemia cells are human leukemia cells.
17. The method according to claim 14 , wherein the leukemia cells are acute myelocytic cells.
18. The method according to claim 14 , wherein the leukemia cells are chronic lymphocytic leukemia cells.
19. The method according to claim 14 , wherein the culture medium comprises exogenous growth factors.
20. The method according to claim 14 , wherein the culture medium includes granulocyte colony stimulating factor, granulocyte monocyte colony stimulating factor, IL3, or IL2.
21. The method according to claim 14 , further comprising:
reseeding the chamber with mononuclear cells.
22. The method according to claim 14 , wherein the mononuclear cells are from peripheral blood, bone marrow aspirate or biopsy, chloroma, or spleen.
23. A method of culturing leukemia cells, comprising:
isolating mononuclear cells, which contain leukemia cells, from a subject and
culturing the cells in a chamber having a scaffolding covered or surrounded with culture medium, wherein said scaffolding is formed from a material selected from the group consisting of metal, glass, ceramic, plastic, hydroxyapatite, a synthetic polymer, a semisynthetic material, and combinations thereof, and the scaffolding allows for leukemia cells to have cell to cell contacts in three dimensions.
24. The method according to claim 23 , wherein the material is degradable.
25. The method according to claim 23 , wherein the material is non-degradable.
26. The method according to claim 23 , wherein the leukemia cells are mammalian leukemia cells.
27. The method according to claim 24 , wherein the mammalian leukemia cells are human leukemia cells.
28. The method according to claim 23 , wherein the leukemia cells are acute myelocytic cells.
29. The method according to claim 23 , wherein the leukemia cells are chronic lymphocytic leukemia cells.
30. The method according to claim 23 , wherein the culture medium comprises exogenous growth factors.
31. The method according to claim 23 , wherein the culture medium includes granulocyte colony stimulating factor, granulocyte monocyte colony stimulating factor, IL3, or IL2.
32. The method according to claim 23 further comprising:
reseeding the chamber with mononuclear cells.
33. The method according to claim 23 , wherein the mononuclear cells are from peripheral blood, bone marrow aspirate or biopsy, chloroma, or spleen.