Compositions and methods of vascular injury repair
The present invention relates to pharmaceutical compositions comprising a chemotactic hematopoietic stem cell product comprising an enriched population of CD34+ cells containing a subpopulation of cells having chemotactic activity, methods of preparing these compositions and use of these compositions to treat or repair vascular injury, including infarcted myocardium.
1. A sterile pharmaceutical composition for repair or a vascular injury caused by vascular insufficiency comprising:
(a) a vascular injury-repairing amount of a sterile chemotactic hematopoietic stem cell product, wherein the vascular injury-repairing amount is effective to repair a vascular injury caused by vascular insufficiency, the chemotactic hematopoietic stem cell product comprising a nonexpanded, isolated population of autologous mononuclear cells enriched for CD34+cells, which further contains a subpopulation of potent CD34+cells that express CXCR-4 and that have CXCR-4-mediated chemotactic activity, and
(b) a stabilizing amount of serum, wherein the stabilizing amount of serum is greater than 20% (v/v),
wherein the pharmaceutical composition is further characterized as having the following properties for at least 24 hours following acquisition of the chemotactic hematopoietic stem cell product when tested in vitro after passage through a catheter:
(i) at least 70% of the cells are CD34+cells,
(ii) contains a subpopulation of potent CD34+/CXCR-4+cells that have CXCR-4-mediated chemotactic activity;
(iii) is at least about 70% viable; and
(iv) is able to form hematopoietic colonies in vitro.
2. The sterile pharmaceutical composition according to claim 1 , wherein the nonexpanded, isolated population of autologous mononuclear cells containing CD34+cells, which further contains a subpopulation of potent CD34+cells that express CXCR-4 and that have CXCR-4-mediated chemotactic activity is purified from cellular components of a bone marrow aspirate.
3. The sterile pharmaceutical composition according to claim 1 , wherein the pharmaceutical composition is further characterized as having the following properties for at least 48 hours following acquisition of the chemotactic hematopoietic stem cell product when tested in vitro after passage through a catheter:
(a) is capable of forming hematopoietic colonies in vitro; and
(b) retains at least 2% of the chemotactic activity of the CXCR-4-mediated chemotactic activity of the subpopulation of potent CD34+/CXCR-4+cells that have CXCR-4-mediated chemotactic activity.
4. The sterile pharmaceutical composition according to claim 1 , wherein the pharmaceutical composition is further characterized as having the following properties for at least 72 hours following acquisition of the chemotactic hematopoietic stem cell product when tested in vitro after passage through a catheter:
(a) is capable of forming hematopoietic colonies in vitro; and
(b) retains at least 2% of the chemotactic activity of the CXCR-4-mediated chemotactic activity of the subpopulation of potent CD34+/CXCR-4+cells that have CXCR-4-mediated chemotactic activity.
5. The sterile pharmaceutical composition of claim 1 , wherein the sterile pharmaceutical composition retains at least 2% of the chemotactic activity-of the CXCR-4-mediated chemotactic activity of the subpopulation of potent CD34+/CXCR-4+cells that have CXCR-4-mediated chemotactic activity for at least 24 hours following acquisition of the chemotactic hematopoietic stem cell product when tested in vitro after passage through a catheter.
6. The sterile pharmaceutical composition according to claim 1 , wherein the composition is formulated for parenteral administration into a coronary blood vessel.
7. The sterile pharmaceutical composition according to claim 1 , wherein the catheter is a flow control catheter.
8. The sterile pharmaceutical composition according to claim 1 , wherein the catheter is a balloon catheter.
9. The sterile pharmaceutical composition according to claim 1 , wherein the catheter has an internal diameter of at least about 0.36 mm.
10. The sterile pharmaceutical composition according to claim 1 wherein the composition is formulated for parenteral administration into myocardium.
11. The sterile pharmaceutical composition according to claim 1 , wherein the pharmaceutical composition further includes at least one compatible active agent.
12. The sterile pharmaceutical composition according to claim 1 , wherein sterility of the chemotactic hematopoietic cell product is confirmed by a method comprising the steps:
(a) centrifuging the chemotactic hematopoietic cell product to form (i) a separated cell product comprising a pellet comprising the -isolated population of mononuclear cells enriched for CD34+cells and (ii) a supernatant;
(b) removing the supernatant of the separated cell product without disturbing the cell pellet of the separated cell product.
(c) analyzing the sterility of the supernatant of the separated cell product, thereby determining the sterility of the cell pellet of the separated cell product without depleting the chemotactic hematopoietic cell product.
13. The sterile pharmaceutical composition according to claim 1 , wherein the nonexpanded, isolated population of autologous mononuclear cells containing CD34+cells, which further contains a subpopulation of potent CD34+cells that express CXCR-4 and that have CXCR-4-mediated chemotactic activity is purified from peripheral blood.
14. The sterile pharmaceutical composition according to claim 1 , wherein the vascular insufficiency results from occlusion of a coronary artery.
15. The sterile pharmaceutical composition according to claim 14 , wherein the vascular insufficiency resulting from occlusion of a coronary artery is a microvascular insufficiency.
16. The sterile pharmaceutical composition according to claim 14 , wherein the vascular insufficiency resulting from occlusion of a coronary artery is an ischemia.
17. The sterile pharmaceutical composition according to claim 14 , wherein the vascular insufficiency resulting from occlusion of a coronary artery is a transient vascular insufficiency.
18. The sterile pharmaceutical composition according to claim 14 , wherein the vascular insufficiency resulting from occlusion of a coronary artery is a myocardial ischemia.
19. The sterile pharmaceutical composition according to claim 14 , wherein the vascular insufficiency resulting from occlusion of a coronary artery produces a myocardial infarction.
20. The sterile pharmaceutical composition according to claim 1 , wherein the vascular injury-repairing amount of the chemotactic hematopoietic stem cell product comprises at least 0.5×10 6 potent CD34+cells that express CXCR-4 and that have CXCR-4 mediated chemotactic activity.
21. The sterile pharmaceutical composition according to claim 1 , wherein the CD34+/CXCR-4+cells of the sterile pharmaceutical composition migrate to and repair damage caused by the vascular insufficiency.
22. The sterile pharmaceutical composition according to claim 21 , wherein migration of the CD34+/CXCR-4+cells of the sterile pharmaceutical composition is driven by chemotaxis.
23. The sterile pharmaceutical composition according to claim 22 , wherein the chemotaxis is Mediated by at least one of SDF-1 and CXCR-4.