Factors and cells that provide for induction of bone, bone marrow, and cartilage
Methods, compositions and kits for producing functional chondrocytes, skeletal cells, bone marrow stromal cells, and progenitor cells thereof are provided. These methods, compositions and kits find use in producing chondrocytes, osteoblasts, stromal cells, and progenitor cells thereof in vivo, or in vitro for transplantation, for experimental evaluation, as a source of lineage- and cell-specific products, and the like, for example for use in treating human disorders of the cartilage, bone and hematopoietic system. In some embodiments, specific combinations of protein factors are identified for reprogramming non-skeletal cells into bones, hematopoietic stroma, and chondrocytes, which may be provided in vitro or in vivo.
1. A method for regenerating articular cartilage in a human individual, the method comprising:
administering to the individual at a site where regeneration of articular cartilage is desired, a combination of an effective dose of human adipose-derived stem cells (hASC) that express BMP receptor (BMPR1); human BMP2 protein in a dose effective to reprogram the ASC to CD45 − CD235 − Tie2 − CD31 − PDPN + CD146 − CD73 + CD164+ human skeletal stem cells (hSSC); and a VEGF inhibitor in a dose effective to induce a chondrogenic fate in the hSSC;
wherein articular cartilage is regenerated at the site.
2. The method of claim 1 , wherein the combination of cells, BMP2 and VEGF inhibitor are provided in a matrix.
3. The method of claim 1 wherein the cells are autologous to the individual.
4. The method of claim 1 wherein the cells are allogeneic to the individual.
5. The method of claim 1 , wherein the cells are freshly isolated from lipoaspirate.
6. The method of claim 2 , wherein the matrix is a scaffold, paste or implant, and wherein articular cartilage is formed at the site of the matrix.
7. The method of claim 1 , wherein the VEGF inhibitor is an antibody that specifically binds to human VEGF.
8. The method of claim 1 , wherein the VEGF inhibitor is a soluble VEGF receptor.
9. The method of claim 1 , wherein the VEGF inhibitor is selected from: ABT-869; AEE-788; AG-13736; AG-028262; Angiostatin; bevacizumab; AVE-8062; AZD-2171; sorafenib; BMS-387032; CEP-7055; CHIR-258; CP-547632; 786034; GW-654652; IMC-1C11; KRN-951; PKC-412; benzoylstaurosporine; CGP-41251; midostaurin; STI-412; PTK-787; vatalanib; sunitinib; semaxanib; SU-666; VEGF Trap; Thalidomide; XL-647; XL-999; XL-880; ZD-6474; and ZK-304709.
10. The method of claim 2 , wherein the matrix is a biodegradable matrix comprised comprising one or more of matrigel, polylactic acid, polyglycolic acid, poly(lactic-co-glycolic acid); collagen, and alginate.
11. The method of claim 5 , wherein the hASC are isolated by selecting for cells that express BMPR1.
12. The method of claim 5 , wherein the hASC are isolated by depleting lipoaspirate of hematopoietic cells.
13. The method of claim 1 wherein following the administering steps, an articular cartilage mass of the treated individual is increased at least 50% relative to an untreated control.
14. A method for regenerating articular cartilage in a human individual, the method comprising:
isolating human adipose stem cells (hASC) from lipoaspirate by negative selection for CD45 and CD235; and positive selection for expression of BMP receptor (BMPR1);
administering to the individual at a site where regeneration of cartilage is desired, a composition consisting of human BMP2, wherein human BMP2 is provided at a concentration from 1 mg/ml to 1 mg/ml; an effective dose of the isolated human adipose-derived stem cells (hASC) that express BMP receptor (BMPR1) to reprogram the ASC to human skeletal stem cells (hSSC), and the VEGF inhibitor is administered in a dose effective to induce a chondrogenic fate in the hSSC;
wherein following the administering steps, the articular cartilage mass is increased at least 50% relative to an untreated control.
15. The method of claim 14 wherein the cells, BMP2 and VEGF inhibitor are provided in a matrix.
16. The method of claim 15 , wherein the matrix is a scaffold, paste or implant, and wherein articular cartilage is formed at the site of the matrix.
17. The method of claim 2 , wherein the matrix is a biodegradable matrix comprised comprising one or more of matrigel, polylactic acid, polyglycolic acid, poly(lactic-co-glycolic acid); collagen, alginate.
18. The method of claim 14 , wherein the VEGF inhibitor is an antibody that specifically binds to human VEGF.
19. The method of claim 14 , wherein the VEGF inhibitor is a soluble VEGF receptor.