Claims
- 1. A suspension of cells comprising pluripotent hematopoietic stem cells substantially free of γc+ cells.
- 2. The cell suspension of claim 1 wherein the pluripotent hematopoietic stem cells comprise single cells having both lymphoid and myeloid potential.
- 3. The cell suspension of claim 2 wherein greater than about 10% of single cells comprise pluripotent hematopoietic stem cells having both lymphoid and myeloid potential.
- 4. The cell suspension of claim 2 wherein greater than about 5% of cells comprise pluripotent hematopoietic stem cells that proliferated late as measured by the Extended Long Term Culture-Initiating Cell (ELTC-IC) assay.
- 5. The cell suspension of claim 1 wherein the pluripotent hematopoietic stem cells comprise CD34+ cells.
- 6. The cell suspension of claim 5 comprising greater than about 85% CD34+ γc− cells.
- 7. The cell suspension of claim 6 comprising greater than about 85% CD34+CD38− γc− cells.
- 8. The cell suspension of claim 6 comprising greater than about 85% CD34+CD38− γc− IL-7Rα− cells.
- 9. A method of purifying hematopoietic stem cells comprising
(a) obtaining a sample of cells that includes hematopoietic stem cells; (b) separating the cells in the sample into CD34+ and CD34− cells; (c) collecting the CD34+ cells; (d) separating the CD34+ cells into CD34+ γc− and CD34+ γc+ cells; (e) collecting the CD34+ γc− cells.
- 10. The method of claim 9 wherein (d) and (e) are modified as follows
(d) separating CD34+ cells into a first group of CD34+ γc− IL-7Rα− cells and a second group of CD34+ γc+ IL-7Rα−, CD34+ γc− IL-7Rα+ and CD34+ γc+ IL-7Rα+ cells; (e) collecting the first group of cells.
- 11. A suspension of human cells from marrow or blood comprising CD34+ γc− cells and substantially free of cells that are not CD34+ γc−, said suspension having the ability to restore the production of lymphoid and hematopoietic cells to a human lacking said production.
- 12. A suspension of normal human cells derived from blood or bone marrow of an individual affected with leukemia comprising normal pluripotent hematopoietic stem cells substantially free of γc+ leukemia cells, mature lymphoid cells and mature myeloid cells.
- 13. The suspension of normal human cells of claim 12, wherein the normal pluripotent hematopoietic stem cells are substantially free of CD34+ γc+ IL-7Rα−, CD34+γc− IL-7Rα+ and CD34+ γc+ IL-7Rα+ leukemia cells, mature lymphoid cells and mature myeloid cells.
- 14. A method of purifying normal pluripotent hematopoietic stem cells substantially free of malignant cells from an individual having a γc+ malignancy comprising
(a) obtaining a sample of cells that includes hematopoietic stem cells from an individual having a γc+ malignancy; (b) separating the cells in the sample into CD34+ and CD34− cells; (c) collecting the CD34+ cells; (d) separating the CD34+ cells into CD34+ γc− and CD34+ γc+ cells; (e) collecting the CD34+ γc− cells.
- 15. The method of claim 14 wherein the individual has a γc+ IL-7Rα+ malignancy, wherein (d) and (e) are modified as follows
(d) separating CD34+ cells into a first group of CD34+ γc− IL-7Rα− cells and a second group of CD34+ γc+ IL-7Rα−, CD34+ γc− IL-7Rα+ and CD34+ γc+ IL-7Rα+ cells; (e) collecting the first group of cells.
- 16. The method of claim 14, wherein the malignancy is leukemia.
- 17. The method of claim 16, wherein the leukemia is selected from the group consisting of acute non-lymphoblastic leukemia (ANLL) or acute lymphoblastic leukemia (ALL).
- 18. A kit for purifying hematopoietic stem cells, said kit having a package comprising
a means for selecting CD34+ cells, said means selected from the group consisting of labeled anti-CD34 antibodies, for labeling cells for separation by a mechanical cell sorter that detects the presence of the label, and anti-CD34 antibodies attached to a first solid support; a means for selecting γc− cells, said means selected from the group consisting of labeled anti-γc antibodies, for labeling cells for separation by a mechanical cell sorter that detects the presence of the label, and anti-γc antibodies attached to a second solid support.
- 19. The kit of claim 18 wherein the first solid support is selected from the group consisting of agarose beads, polystyrene beads, magnetic beads, hollow fiber membranes and plastic petri dish.
- 20. The kit of claim 18 wherein the second solid support is selected from the group consisting of agarose beads, polystyrene beads, magnetic beads, hollow fiber membranes, plastic support and plastic petri dish.
- 21. The kit of claim 18 wherein the anti-CD34 antibodies are labeled with a fluorescent label or a magnetic label.
- 22. The kit of claim 18 wherein the anti-γc antibodies are labeled with a fluorescent label or a magnetic label.
- 23. The kit of claim 18 wherein the anti-γc antibodies are monoclonal antibodies.
- 24. The kit of claim 23 wherein the anti-γc antibodies are γy1 monoclonal antibodies.
- 25. A kit for purifying normal pluripotent hematopoietic stem cells substantially free of malignant cells from an individual having a γc+ malignancy comprising, said kit having a package comprising
a means for selecting CD34+ cells, said means selected from the group consisting of labeled anti-CD34 antibodies, for labeling cells for separation by a mechanical cell sorter that detects the presence of the label, and anti-CD34 antibodies attached to a first solid support; a means for selecting γc− cells, said means selected from the group consisting of labeled anti-γc antibodies, for labeling cells for separation by a mechanical cell sorter that detects the presence of the label, and anti-γc antibodies attached to a second solid support.
- 26. The kit of claim 25 wherein the first solid support is selected from the group consisting of agarose beads, polystyrene beads, magnetic beads, hollow fiber membranes and plastic petri dish.
- 27. The kit of claim 25 wherein the second solid support is selected from the group consisting of agarose beads, polystyrene beads, magnetic beads, hollow fiber membranes, plastic support and plastic petri dish.
- 28. The kit of claim 25 wherein the anti-CD34 antibodies are labeled with a fluorescent label or a magnetic label.
- 29. The kit of claim 25 wherein the anti-γc antibodies are labeled with a fluorescent label or a magnetic label.
- 30. The kit of claim 25 wherein the anti-γc antibodies are monoclonal antibodies.
- 31. The kit of claim 30 wherein the anti-γc antibodies are γc1 monoclonal antibodies.
- 32. A method of reconstituting the lympho-hematopoietic system of an individual in need thereof comprising administering an effective amount of the cell suspension of claim 1.
- 33. A method of reconstituting the lympho-hematopoietic system of an individual in need thereof comprising administering an effective amount of the cell suspension of claim 12.
RELATED APPLICATIONS
[0001] This application claims priority to the U.S. Provisional Application Serial No. 60/162,348, filed Oct. 29, 1999, by Crooks, et al., and entitled “γc Negative Hematopoietic Progenitor Cells,” which is incorporated herein by reference in its entirety, including any drawings.
GOVERNMENT LICENSE RIGHTS
[0002] The U.S. Government has a paid-up license in this invention and the right in limited circumstances to require the patent owner to license others on reasonable terms as provided for by the terms of Grant No. A140581 awarded by NIH.
Provisional Applications (1)
|
Number |
Date |
Country |
|
60162348 |
Oct 1999 |
US |
Continuations (1)
|
Number |
Date |
Country |
Parent |
PCT/US00/29143 |
Oct 2000 |
US |
Child |
10136855 |
Apr 2002 |
US |