Enhancement of angiogenesis to grafts using cells engineered to produce growth factors
The present invention provides methods and compositions of engineered cells for use in the continuous or transient delivery of growth factors and angiogenesis modulating agents, such as vascular endothelial growth factor (VEGF), in conjunction with constructs for replacing or augmenting organ functions. In one aspect of the invention, the genetically engineered cells can be immature cells that are capable of differentiating and assimilating into the target region. The methods of the present invention can be used to enhance vascularization locally at a target site in need of repair, growth, or implantation through the incorporation of autologous cells which have been genetically engineered to secrete a growth factor or angiogenesis modulating agent.
1. A method for augmenting organ function comprising:
transiently transfecting a first population of cells with a plasmid encoding an angiogenesis modulating agent;
culturing at least a second population of cells on a matrix material to produce an organ construct, wherein the second population of cells comprises cells of a different cell type than the first population; and
implanting the organ construct and the first population of cells in vivo at a target site to replace or augment organ function, such that the first population of cells express the angiogenesis modulating agent thereby inducing the second population of cells to assimilate and differentiate at the target site.
2. The method of claim 1 , wherein the matrix is decellularized tissue.
3. The method of claim 1 , wherein the matrix is a hydrogel.
4. The method of claim 1 , Wherein the matrix is a polymer.
5. The method of claim 1 , Wherein the angiogenesis modulating agent is VEGF.
6. The method of claim 1 , wherein the method further comprises assimilating the first population of cells into a tissue layer.
7. The method of claim 1 , wherein the step of transiently transfecting the first population of cells further comprises:
encapsulating the transfected first population of cells and
implanting the organ construct and the encapsulated first population of cells in vivo at the target site to replace or augment organ function such that the first population of cells express the angiogenesis modulating agent and the second population of cells assimilate and differentiate at the target site.
8. The method of claim 7 , wherein the step of encapsulating the transfected first population of cells further comprises using microspheres.
9. The method of claim 7 , wherein the step of encapsulating the transfected first population of cells further comprises using alginate-PLL capsules.
10. The method of claim 1 , wherein the first population of cells comprises endothelial progenitor cells.
11. The method of claim 1 , wherein the first population of cells comprises vascular endothelial cells (EC).
12. The method of claim 1 , wherein the first population of cells comprises myoblasts.
13. The method of claim 1 , wherein the second population of cells comprises endothelial progenitor cells.
14. The method of claim 1 , wherein the second population of cells comprises myoblasts.
15. A method of organ augmentation comprising the steps of:
transiently transfecting a first population of cells with a plasmid encoding the angiogenesis modulating agent VEGF;
selecting a second population of cells to be assimilated at a target tissue region upon implantation,
suspending the first population of cells and the second population of cells in an injectable polymer matrix;
injecting the first population of cells and the second population of cells and the polymer matrix into the target tissue region where the first population of cells will express the VEGF angiogenesis modulating agent, thereby inducing assimilation and differentiation of the second population of cells in the target region and augmenting organ function.
16. The method of claim 15 , wherein the polymer matrix comprises collagen.
17. The method of claim 16 , wherein the polymer matrix comprises collagen type I.
18. The method of claim 1 , wherein the step of transiently transfecting the first population of cells further comprises:
encapsulating the transfected first population of cells;
suspending the encapsulated first population of cells and the second population of cells in an injectable polymer matrix
injecting the encapsulated first population of cells and the second population of cells and the polymer matrix into the target tissue region where the encapsulated first population of cells will express the VEGF angiogenesis modulating agent, thereby inducing assimilation and differentiation of the second population of cells in the target region and augmenting organ function.
19. The method of claim 18 , wherein the step of encapsulating the transfected first population of cells further comprises using microspheres.
20. The method of claim 18 , wherein the step of encapsulating the transfected first population of cells further comprises using alginate-PLL capsules.
21. The method of claim 15 , wherein the step of transfecting the first population of cells comprises transiently transfecting the cells such that the angiogenesis modulating agent is produced for less than three weeks.
22. The method of claim 15 , wherein the first population of cells comprises undifferentiated cells.
23. The method of claim 15 , wherein the first population of cells comprises endothelial progenitor cells.
24. The method of claim 15 , wherein the first population of cells comprises vascular endothelial cells (EC).
25. The method of claim 15 , wherein the first population of cells comprises myoblasts.
26. The method of claim 15 , wherein the second population of cells comprises endothelial progenitor cells (EPC).
27. The method of claim 1 , wherein the second population of cells comprises myoblasts.