Microcapsule cross-linking

Abstract

A method for the gentle formation of microcapsule populations is disclosed. Capsule encapsulation is achieved using a hydrogel reagent that reduces or eliminates the negative effects of hydrogel crosslinking in the microcapsule contents while enabling the release of the contents under physiological conditions. Also disclosed are the resulting microcapsule populations that contain contents such as various cell populations in proportions that reflect their relative proportions before encapsulation, so that encapsulation does not differentially select one or another cell population.

Claims

1. A microcapsule population comprising at least 100 microcapsules, wherein each microcapsule comprises a cross-linked shell and an aqueous interior, and the aqueous interior contains living cells such that the microcapsule population comprises at least 100 cells.

2. The microcapsule population according to claim 1, wherein certain parameters of the cells in the microcapsule population differ from those of the corresponding cells in the microcapsule precursor sample population by no more than 10%.

3. The microcapsule population according to claim 2, wherein the parameters include a transcription accumulation pattern.

4. The microcapsule population according to claim 2, wherein the parameters include a proteome accumulation pattern.

5. The microcapsule population according to claim 2, wherein the parameter includes growth rate.

6. The microcapsule population according to claim 2, wherein the parameter includes cell viability dye fluorescence.

7. The microcapsule population according to claim 2, wherein the parameter includes cell motility.

8. The microcapsule population according to claim 1, wherein the proportional cell identity of at least 100 capsule-encapsulated cells differs from a population of cells selected from a microcapsule sample precursor population by no more than 10%.

9. The microcapsule cluster according to claim 1, wherein the microcapsule cluster is degradable under physiological conditions.

10. The microcapsule cluster according to claim 1, wherein the microcapsule cluster is enzymatically degradable.

11. The microcapsule assembly according to claim 1, wherein the microcapsule assembly is degradable by treatment with dextranase.

12. The microcapsule population according to claim 1, wherein at least a portion of living cells proliferate and form colonies within the microcapsules of the population.

13. A method for forming a microcapsule population, each comprising microcapsules having a cross-linked shell and an aqueous interior containing living cells, the method comprising the step of preventing oxidative stress from being active in the precursor of the liquid core of the microcapsules of the population.

14. The method according to claim 13, wherein forming the composition includes the step of expelling dissolved oxygen molecules from the internal space of the capsule.

15. The method according to claim 14, wherein the expulsion step includes flushing the internal space with an oxygen-removing gas.

16. The method according to claim 13, wherein forming the composition comprises the step of contacting an aqueous emulsion with a carrier, the oxidative stress portion comprising a non-water-soluble crosslinking initiator in the carrier.

17. The method according to claim 16, wherein the non-water-soluble crosslinking initiator is present in an insufficient concentration to allow the radical to reach the internal space of the capsule.

18. The method according to claim 13, comprising the step of forming a composition which prevents free radical stress from being active in the precursor of the liquid core of the collective microcapsules.

19. The method according to claim 18, wherein forming the composition comprises the step of contacting an aqueous emulsion with a carrier having a free radical stress-inducing moiety, the free radical stress moiety containing a water-insoluble crosslinking initiator in the carrier.

20. The method according to claim 19, wherein the non-water-soluble crosslinking initiator is present in an insufficient concentration to allow the radical to reach the internal space of the capsule.

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