Particulate delivery vehicles for embryoid bodies
The present invention provides a vehicle for delivering various chemicals, compositions and proteins to stem cells and embryoid bodies. The vehicle may be biocompatible and biodegradable polymer microparticles. Typically the particles will contain at least a growth factor for delivery to the embryoid bodies, and generally the growth factor induces differentiation of the cells in the embryoid body along a specific lineage. The present invention also provides methods for directing differentiation of the cells in the embryoid body.
1. A method of directing differentiation of stem cells comprising the steps of:
providing human embryonic stem cells;
providing microparticles with a diameter in the range of between about 0.2 μm and about 6 μm, wherein the microparticles comprise a growth factor;
contacting the embryonic stem cells with the microparticles, in a solution, under conditions suitable to form aggregates comprising the embryonic stem cells and the microparticles and to affect differentiation of the cells in the formation of the embryoid bodies.
2. The method of claim 1 wherein the contacting step comprises centrifuging the mixture of cells and microparticles.
3. The method of claim 1 wherein the microparticles are comprised of a bio-compatible and biodegradable polymer.
4. The method of claim 3 wherein the polymer comprises poly(lactide-co-glycolide).
5. The method of claim 3 wherein the microparticles are comprised of a material selected from the group comprising poly(anhydrides), poly(hydroxy acids), poly(ortho esters), poly(propylfumerates), poly(caprolactones), polyamides, polyamino acids, polyacetals, biodegradable polycyanoacrylates, biodegradable polyurethanes, poly(glycerol sebacates), elastomeric poly(glycerol sebacates polysaccharides), and combinations thereof.
6. The method of claim 1 wherein the microparticles are comprised of a material selected from the group comprising polypyrrole, polyanilines, polythiophene, polystyrene, polyesters, non-biodegradable polyurethanes, polyureas, poly(ethylene vinyl acetate), polypropylene, polymethacrylate, polyethylene, polycarbonates, poly(ethylene oxide), co-polymers and combinations thereof.
7. The method of claim 1 wherein the microparticles comprise VEGF165 as the growth factor and the cells, after contacting with the microparticles, express CD 34.
8. The method of claim 1 wherein the microparticles comprise PlGF and/or bFGF as the growth factor and the cells, after contacting with the microparticles, express PECAM1.
9. The method of claim 1 wherein the growth factor is selected from the group consisting of vascular endothelial growth factor (VEGF165), basic fibroblast growth factor (bFGF), placenta growth factor (PlGF), and combinations thereof.
10. The method of claim 1 wherein the growth factor is selected from the group comprising epidermal growth factor, nerve growth factor, transforming growth factor-β, platelet-derived growth factor, insulin-like growth factor, acidic fibroblast growth factor, basic fibroblast growth factor, hepatocyte growth factor, brain-derived neurotrophic factor, keratinocyte growth factor, bone morphogenetic protein, a cartilage-derived growth factor, and combinations thereof.
11. The method of claim 1 wherein the microparticles may be taken up by the cells of the embryoid bodies.
12. The method of claim 1 wherein the first providing step comprises providing about 30,000 stem cells.