Microencapsulation of living tissue and cells

• US 4,673,566 A
• Filed: 05/24/1984
• Issued: 06/16/1987
• Est. Priority Date: 06/01/1983
• Status: Expired due to Fees

• Current Assignee: Connaught Laboratories Limited (Sanofi )
• Original Assignee: Connaught Laboratories Limited (Sanofi )
• Inventors: Goosen, Mattheus F. A., Sun, Anthony M., O'Shea, Geraldine M.
• Primary Examiner(s): Rosen, Sam
• Application Number:
  US06/613,780
• Time in Patent Office: 1,118 Days
• Field of Search: 424/95, 424/110, 424/21, 424/16, 424/19, 424/31, 424/32, 424/35, 424/93, 435/240, 435/241, 435/1, 604/890, 604/891
• US Class Current: 424/424
• CPC Class Codes: A61K 9/1652 Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin homeopathic globules A61K9/1623
• CPC Class Codes: A61K 9/5031 obtained otherwise than by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyethylene glycol, poly(lactide-co-glycolide)
• CPC Class Codes: A61K 9/5036 Polysaccharides, e.g. gums, alginate; Cyclodextrin
• CPC Class Codes: A61K 9/5073 having two or more different coatings optionally including drug-containing subcoatings
• CPC Class Codes: Y10S 930/29 Polyamino acid or polypeptide with an uninterrupted series of peptide repeating units

Abstract

Living tissue or cells, for example, islets of Langerhans, are microencapsulated for implantation in the body for long term treatment of diabetes or other disease requiring organ transplantation. The microcapsules take the form of a biocompatible semi-permeable hydrogel membrane based on polylysine which permits the passage of materials and oxygen to the cells and metabolic products from the cells while retaining the cells encapsulated. The biocompatible semi-permeable membrane has an outer negatively-charged surface, which, combined with a controlled thickness of polylysine of molecular weight from 10,000 to 30,000 daltons, imparts to the microcapsules the ability to maintain long term effectiveness.

Claims

1. A microcapsule having a diameter of about 500 to about 2000 μ

m and suitable for implantation into an animal body, comprising;

a core comprising one or more viable, healthy, physiologically-active tissue cells capable of ongoing metabolism, and

a biocompatible semi-permeable membrane surrounding and enclosing said core, said semi-permeable membrane being permeable to tissue nutrients and metabolic products produced by the tissue but impermeable to immune system proteins, said semi-permeable membrane having a molecular weight cut-off of below about 150,000 daltons,

said biocompatible membrane being a hydrogel formed by ionic reaction between a polylysine polymer having a molecular weight from about 10,000 to about 30,000 daltons and a polymeric material bearing negatively-charged groups to provide an outer negatively-charged surface,

said polylysine polymer membrane having a durability sufficient to permit said microcapsules to be injected in the animal body and sufficient to maintain said microcapsules in an intact form and to permit said tissue to effect ongoing metabolism when injected into the animal body for a period of time exceeding three months.

2. The microcapsule of claim 1 wherein said tissue cells are islets of Langerhans.

3. The microcapsule of claim 1 wherein said negatively charged groups are carboxyl or hydroxyl groups.

4. The microcapsule of claim 3 wherein said polymeric material bearing negatively charged groups is selected from the group consisting of alginate, polyvinyl alcohol or polylactic acid.

5. The microcapsule of claim 4 wherein said polylysine has a molecular weight of about 15,000 to about 25,000.

6. The microcapsule of claim 5 wherein said polylysine has a molecular weight of about 17,000.

7. The microcapsule of claim 1 having a diameter of about 700 to about 1000 μ

m and wherein said semi-permeable membrane has a thickness of about 5 μ

m.

8. The microcapsule of claim 1 wherein said membrane has a molecular weight cut-off of about 67,000 daltons.

9. The microcapsule of claim 1 wherein said core also comprises an aqueous medium of nutrients sufficient to maintain said tissue cells and allow normal metabolism thereof.

10. A method of encapsulating a core material within a semi-permeable membrane to form microcapsules for implantation into an animal body, which comprises:

(a) placing the material in an aqueous solution of a water-soluble polymeric substance that can be reversibly gelled and which has free acid groups,

(b) forming the solution into droplets,

(c) gelling the droplets to produce discrete shape-retaining temporary capsules,

(d) forming semi-permeable membranes about the temporary capsules by contact between the temporary capsules and a polymer containing free amino groups to cause ionic reaction with the acid groups in a surface layer of the capsule, said polymer containing free amino groups being polylysine having a molecular weight of about 10,000 to about 30,000 daltons, said contact being effected for a period of time no less than about four minutes sufficient to provide a polymer coating on the temporary capsule of sufficient durability to permit the microcapsules to be injected into the animal body, and

(e) contacting said microcapsules formed in step (d) with a biocompatible polymeric material which contains free negatively-charged groups capable of ionic reaction with the free amino groups in a surface layer of the microcapsule, thereby to form an outer coating of said biocompatible polymeric material on said microcapsules.

11. The method of claim 10 wherein said core material comprises living tissue which is in finely-divided suspended form in said aqueous solution in step (a).

12. The method of claim 11 wherein said living tissue comprises islets of Langerhans whereby said microcapsules may be used to control blood sugar levels in diabetic animal bodies into which the microcapsules are implanted.

13. The method of claim 10 wherein said reversibly-gellable water-soluble substance is a polysaccharide gum.

14. The method of claim 13 wherein said gum is an alkali metal alginate.

15. The method of claim 10 wherein said polylysine has a molecular weight of about 15,000 to about 25,000 daltons.

16. The method of claim 10 wherein said contact in step (d) is effected for a period of time sufficient to provide a polylysine layer of thickness about 5 μ

m.

17. The method of claim 10 wherein said contact in step (d) is effected by contact with an aqueous solution of polylysine at about 6 to about 9 minutes.

18. The method of claim 17, wherein said aqueous solution of polylysine has a concentration of at least about 0.05 wt.%.

19. The method of claim 10 wherein said biocompatible negatively-charged polymeric material is selected from the group consisting of polyvinyl alcohols having free hydroxyl groups and polylactic acids containing free acid groups.

20. The method of claim 10 wherein said reversibly-gellable water-soluble substance comprises sodium alginate, and said biocompatible polymeric material comprises sodium alginate.

21. The method of claim 14 wherein said alkali metal alginate is sodium alginate and the viscosity of said aqueous solution of sodium alginate is at least sufficient to result in the formation of substantially spherical temporary capsules.

22. The method of claim 21 wherein said aqueous sodium alginate solution has a viscosity of at least about 30 cps.

23. The method of claim 10 including the further step of reliquifying the gel within the semi-permeable membrane.

24. A tissue implantation method, which comprises:

encapsulating tissue within a spherical microcapsule having a diameter of about 500 to about 2000 μ

m and a biocompatible semipermeable membrane having a negatively-charged biocompatible outer surface and a thickness of about 5 microns, said biocompatible membrane having a molecular weight cut-off below 150,000 daltons and being impermeable to immune system proteins but permeable to tissue nutrients and metabolic products produced by the tissue, said biocompatible semipermeable membrane being a hydrogel formed by ionic reaction between a polylysine polymer having a molecular weight from about 10,000 to about 30,000 daltons and a polymeric material being negatively charged groups and further comprising an outer, negatively charged surface and

introducing said microcapsule into a mammalian body to effect ongoing metabolism in said body for a period of time exceeding three months.

25. The method of claim 24 wherein said microcapsule is introduced by injection.

26. The method of claim 24 wherein said tissue comprises islets of Langerhans or a fraction thereof.

27. The method of claim 24 wherein said microcapsule is substantially perfectly spherical.

28. The method of claim 24 wherein said tissue comprises liver cells.

29. The method of claim 24 wherein said membrane has a molecular weight cut-off of less than about 67,000 daltons.