Claims
- 1. An ex-vivo method of treating myeloablation of hematopoietic stem cells and progenitor cells in a subject, comprising the steps of:
a. isolating said hematopoietic stem and progenitor cells from the bone marrow of a donor; b. expanding said isolated stem and progenitor cells in a co-culture medium including endothelial cells; c. harvesting said expanded stem and progenitor cells from said co-culture medium; d. administering a therapeutic dose of said harvested expanded stem and progenitor cells to said myeloablated subject.
- 2. The method of claim 1, wherein the cause of said myeloablation in said subject is selected from the group consisting of: ionizing radiation, toxins, chemicals, drugs, disease, and genetic disorders.
- 3. The method of claim 2, wherein said hematopoietic cells are isolated from said donor or said subject within 24 hours after said myeloablation.
- 4. The method of claim 1, wherein said culture medium includes at least one cytokine.
- 5. The method of claim 4, wherein said at least one cytokine is selected from the group consisting of granulocyte-macrophage colony stimulating factor, interleukin-3, stem cell factor and interleukin-6, flt3-ligand, and mixtures thereof.
- 6. The method of claim 1, wherein said subject is a mammal.
- 7. The method of claim 1, wherein said medium comprises a monolayer of endothelial cells.
- 8. The method of claim 7, wherein said endothelial cells are porcine microvascular endothelial cells.
- 9. The method of claim 7, wherein said endothelial cells are human brain endothelial cells.
- 10. The method of claim 1, wherein said hematopoietic stem cells are CD34+38−.
- 11. The method of claim 1, wherein said hematopoietic stem cells are harvested after said expansion and before said cells differentiate.
- 12. The method of claim 11, wherein said harvesting occurs after 7 to 14 days of co-culture.
- 13. The method of claim 11, wherein said harvesting occurs after 10 to 14 days of co-culture.
- 14. The method of claim 1, wherein said culture media is maintained at a pH of about 7.2 to about 7.5 during said expanding step.
- 15. The method of claim 14, wherein said pH is maintained by replacing a portion of said culture medium with fresh culture medium.
- 16. The method of claim 1, wherein said therapeutic dose is about 1 to about 2 million cells/kg subject mass.
- 17. The method of claim 16, wherein said therapeutic dose is administered by injection.
- 18. The method of claim 17, wherein said therapeutic injection is administered by venous injection.
- 19. The method of claim 1, wherein said stem cells are CD34+.
- 20. The method of claim 1, wherein said method results in no engraftment defects in said co-cultured cells.
- 21. The method of claim 20, wherein said method results in no differentiation of the hematopoietic stem cell population.
- 22. The method of claim 21, wherein said method results in no alterations in adhesion molecules and homing receptors on said expanded hematopoietic stem cells.
- 23. The method of claim 1, wherein said donor and said subject are autologous.
- 24. The method of claim 1, wherein said donor and said subject are heterologous.
- 25. The method of claim 24, wherein said donor is myeloablated.
- 26. The method of claim 1, wherein said hematopoietic stem cells are enriched at least 85%.
- 27. The method of claim 1, wherein said hematopoietic stem cells are enriched at least 99%.
- 28. An ex-vivo method of restoring a depleted population of rapidly proliferating hematopoietic stem cells, comprising the steps of:
a. isolating hematopoietic stem cells from a donor; b. expanding said isolated hematopoietic stem cells in a co-culture medium including endothelial cells; c. harvesting said expanded hematopoietic stem cells from said co-culture medium; d. administering a therapeutic dose of said harvested expanded hematopoietic stem cells to a subject.
CROSS-REFERENCE TO RELATED APPLICATIONS
[0001] This application claims priority to U.S. Provisional Application 60/330,754, filed Oct. 30, 2001, entitled EX VIVO RESCUE OF TRANSPLANTABLE HEMATOPOIETIC STEM CELLS FOLLOWING HIGH DOSE RADIATION INJURY. This application hereby incorporates by reference and in its entirety, U.S. patent application Ser. No. 09/452,855, filed Dec. 3, 1999, entitled HUMAN BRAIN ENDOTHELIAL CELLS AND GROWTH MEDIUM AND METHOD FOR EXPANSION OF PRIMITIVE CD34+CD38− BONE MARROW STEM CELLS, by Chute et al. as well as references 1 through 36 cited and listed in this application.
Provisional Applications (1)
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Number |
Date |
Country |
|
60330754 |
Oct 2001 |
US |