Methods for purifying hematopoietic stem cells
Described herein are methods for purifying hematopoietic stem cells. Also described herein are methods for purifying EPCR+ cells. The invention also provides substantially pure isolated hematopoietic stems cells, including EPCR+ hematopoietic stem cells.
1. A method for obtaining a substantially pure population of hematopoietic stem cells, comprising:
(a) contacting a biological sample comprising hematopoietic stem cells with an affinity agent which binds to endothelial protein C receptor (EPCR) under conditions appropriate for binding to occur; and
(b) separating cells that bind to the affinity agent from cells that do not bind to the affinity agent, thereby producing a substantially pure population of hematopoietic stem cells.
2. The method of claim 1 , wherein the affinity agent is an antibody or fragment thereof.
3. The method of claim 1 or 2 , wherein the step of separating cells is performed by a method selected from the group consisting of: column chromatography, fluorescence-activated cell sorting, magnetic bead separation and direct immune adherence.
4. The method of claim 1 , wherein the biological sample comprising cells is selected from the group consisting of: bone marrow cells, embryonic yolk sac, fetal liver, fetal and adult spleen and blood.
5. A method for obtaining a substantially pure population of EPCR+ hematopoietic stem cells, comprising:
(a) contacting a biological sample comprising EPCR+ hematopoietic stem cells with an affinity agent which binds to the endothelial protein C receptor (EPCR) under conditions appropriate for binding to occur; and
(b) separating cells that bind to the affinity agent from cells that do not bind to the affinity agent, thereby producing substantially pure population of EPCR+ hematopoietic stem cells.
6. The method of claim 5 , wherein the affinity agent is an antibody or fragment thereof.
7. The method of claim 5 or 6 , wherein the step of separating cells is performed by a method selected from the group consisting of: column chromatography, fluorescence-activated cell sorting, magnetic bead separation and direct immune adherence.
8. The method of claim 5 , wherein the biological sample comprising cells is selected from the group consisting of: bone marrow cells, embryonic yolk sac, fetal liver, fetal and adult spleen and blood.
9. The method of claim 8 , wherein the EPCR+ cells are human EPCR+ cells.
10. The method of claim 8 , wherein the EPCR+ cells are murine EPCR+ cells.