Method for encapsulation of cellular material using a charged initiator polymer
View Patent ↗Positively-charged initiator polymers having a polymerization initiator group and a cationic portion are provided. The initiator polymers can be used with a polymerizable material for the formation of a polymeric matrix on a surface. The initiator polymers are particularly useful for cell encapsulation using macromers.
1. A method for the encapsulation of cellular material comprising the steps of:
(a) providing cellular material comprising cells;
(b) contacting the cellular material with a positively-charged initiator polymer having a polymeric backbone comprising:
(i) at least 5 initiator groups pendent from the polymeric backbone, and
(ii) a cationic portion comprising cationic groups pendent from the polymeric backbone, wherein the cationic groups provide a positive-charge to the positively-charged initiator polymer in conditions having a pH up to and including, pH 8,
wherein the initiator polymer has a ratio of cationic groups:initiator groups of at least 1:1 and has a molecular weight of 250,000 Da or less, and
wherein the cationic groups of the cationic portion interact with negatively charged surface molecules on the cellular material to localize the initiator polymer on the surface of the cellular material;
(c) contacting the cellular material with a polymerizable material; and
(d) activating the initiator groups of the positively-charged initiator polymer to cause polymerization of the polymerizable material and formation of a polymeric layer around the cellular material.
2. The method of claim 1 wherein the positively-charged initiator polymer has a polymeric backbone that comprises nitrogen.
3. The method of claim 2 wherein the polymeric backbone comprises a polyimine backbone.
4. The method of claim 3 wherein the polymeric backbone comprises polyethylenimine.
5. The method of claim 1 wherein the cationic groups comprise a group selected from the group consisting of quaternary ammonium, quaternary phosphonium, and ternary sulfonium.
6. The method of claim 5 wherein the cationic groups comprise a quaternary ammonium group.
7. The method of claim 1 wherein the cationic groups provide a positive charge to the positively-charged initiator polymer in conditions having a pH up to, and including, pH 10.
8. The method of claim 7 wherein the cationic groups provide a positive charge to the positively-charged initiator polymer in conditions having a pH up to, and including, pH 12.
9. The method of claim 1 wherein the initiator groups comprise a photoinitiator.
10. The method of claim 9 wherein the photoinitiator has an excitation wavelength of 350 nm or greater.
11. The method of claim 10 wherein the photoinitiator has an excitation wavelength of 500 nm or greater.
12. The method of claim 11 wherein the photoinitiator is selected from the group consisting of acridine orange, camphorquinone, ethyl eosin, eosin Y, erythrosine, fluorescein, methylene green, methylene blue, phloxime, riboflavin, rose bengal, thionine, and xanthine dyes.
13. The method of claim 1 wherein the positively-charged initiator polymer has a molecular weight in the range of 8,500 to 250,000 Da.
14. The method of claim 13 wherein the positively-charged initiator polymer has a molecular weight in the range of 8,500 to 100,000 Da.
15. The method of claim 1 wherein the positively-charged initiator polymer comprises up to 1000 monomeric units.
16. The method of claim 1 which provides a cell, a group of cells, or a tissue that is encapsulated with said polymeric layer.
17. The method of claim 1 wherein the cellular material comprises a pancreatic islet cell.
18. The method of claim 16 , comprising a step of (e) introducing the cell, the group of cells, or the tissue that is encapsulated with said polymeric layer into a subject.
19. The method of claim 1 wherein the initiator polymer comprises monomeric units in the polymeric backbone, and 20% or more of the monomeric units are coupled to the cationic groups.
20. The method of claim 19 wherein 50% or more of the monomeric units are coupled to the cationic groups.
21. The method of claim 1 wherein the initiator polymer has a ratio of cationic groups:initiator groups of at least 2.5:1.
22. The method of claim 21 wherein the initiator polymer has a ratio of cationic groups:initiator groups of at least 5:1.
23. A method for the encapsulation of cellular material comprising the steps of:
(a) providing a cellular material comprising cells;
(b) contacting the cellular material with a composition comprising a
a polymerizable material, and
a positively-charged initiator polymer having a polymeric backbone comprising:
(i) at least 5 initiator groups pendent from the polymeric backbone, and
(ii) a cationic portion comprising cationic groups pendent from the polymeric backbone, wherein the cationic groups provide a positive-charge to the positively-charged initiator polymer in conditions having a pH up to, and including, pH 8,
wherein the initiator polymer has a ratio of cationic groups:initiator groups of at least 1:1 and has a molecular weight of 250,000 Da or less, and
wherein the cationic groups of the cationic portion interact with negatively charged surface molecules on the cellular material to localize the initiator polymer on the surface of the cellular material; and
(c) activating the initiator groups of the positively-charged initiator polymer to cause polymerization of the polymerizable material and formation of a polymeric layer around the cellular material.
24. The method of claim 1 wherein the polymerizable material comprises a macromer.
25. The method of claim ≧wherein the polymerizable material comprises a polysaccharide macromer or a protein macromer.
26. The method of claim 24 wherein the polymerizable material comprises a macromer selected from the group consisting of poly(vinylpyrrolidone) (PVP) macromers, poly(ethylene glycol) (PEG) macromers, poly(ethylene oxide) poly(ethyloxazoline) macromers, poly(propylene oxide) macromers, polyacrylamide (PAA) macromers, poly(vinyl alcohol) (PVA) macromers, and copolymers thereof.
27. A method for the encapsulation of cellular material comprising the steps of:
(a) providing cellular material comprising cells;
(b) contacting the cellular material with a positively-charged initiator polymer having a polymeric backbone comprising polyethylenimine comprising:
(i) at least 5 initiator groups comprising an eosin initiator group pendent from the polymeric backbone, and
(ii) a cationic portion comprising cationic groups pendent from the polymeric backbone, wherein the cationic groups provide a positive-charge to the positively-charged initiator polymer in conditions having a pH up to, and including, pH 8,
wherein the initiator polymer has a ratio of cationic groups:initiator groups of at least 1:1 and has a molecular weight of 250,000 Da or less, and
wherein the cationic groups of the cationic portion interact with negatively charged surface molecules on the cellular material to localize the initiator polymer on the surface of the cellular material; and
(c) contacting the cellular material with polymerizable material comprising poly(ethylene glycol) (PEG) comprising one or more acrylate groups; and
(d) activating the initiator groups of the positively-charged initiator polymer to cause polymerization of the polymerizable material and formation of a polymeric layer around the cellular material.