Microencapsulation of living tissue and cells

• US 4,806,355 A
• Filed: 06/08/1987
• Issued: 02/21/1989
• Est. Priority Date: 06/06/1983
• Status: Expired due to Term

• Current Assignee: Connaught Laboratories Limited (Sanofi )
• Original Assignee: Connaught Laboratories Limited (Sanofi )
• Inventors: Sun, Anthony M. F., O'Shea, Geraldine M., Goosen, Mattheus F. A.
• Primary Examiner(s): Rosen, Sam
• Application Number:
  US07/059,513
• Time in Patent Office: 624 Days
• Field of Search: 424/16, 424/21, 424/422, 424/423, 424/424, 514/964, 435/240.22
• US Class Current: 424/424
• CPC Class Codes: A61F 2/022 Artificial gland structures using bioreactors hemodialysis A61M1/16; hemofiltration A61M1/34; other extracorporeal blood treatment A61M1/36
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 35/407 Liver; Hepatocytes
• CPC Class Codes: A61K 9/1652 Polysaccharides, e.g. alginate, cellulose derivatives; Cyclodextrin homeopathic globules A61K9/1623
• CPC Class Codes: A61K 9/5026 obtained by reactions only involving carbon-to-carbon unsaturated bonds, e.g. polyvinyl pyrrolidone, poly(meth)acrylates
• 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 9/5073 having two or more different coatings optionally including drug-containing subcoatings
• CPC Class Codes: A61L 27/20 Polysaccharides
• CPC Class Codes: A61L 27/38 containing added animal cells organs or tissue containing native cells A61L27/36
• CPC Class Codes: C12N 11/04 entrapped within the carrier, e.g. gel or hollow fibres
• CPC Class Codes: G01N 15/149 specially adapted for sorting particles, e.g. by their size or optical properties

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 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 imparts to the microcapsules the ability to maintain long term effectiveness.

Claims

1. A biocompatible in microcapsule, suitable for implantation into an animal body and having a diameter of about 50 to about 2000 μ

m, comprising;

a macromolecular spherical core containing living tissue or individual cells thereof, said core being surrounded by of interpenetrating layers of ionically-interacted biocompatible materials defining a membrane thickness of about 5 to about 20 μ

m, said biocompatible materials comprising a biocompatible polyamino acid polymer normally having a positively-charged surface and a non-toxic biocompatible water-soluble polymeric material which is capable of ionic interaction with free amino groups in said polyamino acid to provide an outer biocompatible negatively-charged surface, said biocompatible semi-permeable membrane being in the form of a hydrogel having an overall water content within the membrane structure of at least about 20 wt. %,

said biocompatible semi-permeable membrane being permeable to and permitting nutrients and oxygen to flow from a body in which the microcapsule is implanted to said living tissue of individual cells thereof and permitting metabolic products of said living tissue to flow therefrom to the body in which the microcapsule is implanted and being impermeable to said living tissue to retain the living tissue within the microcapsule, said microcapsule being capable of resisting degradation and remaining permeable in vivo for at least two months.

2. The microcapsules of claim 1 wherein said living tissue is an animal tissue selected from the group consisting of islets of Langerhans, liver and individual cells thereof.

3. The microcapsules of claim 1 wherein said polyamino acid is comprises of polylysine, the inner surface is comprised of alginate and the outer surface is comprised of alginate, polyvinyl alcohol or polylactic acid.

4. The microcapsules of claim 1 wherein said living tissue is islets of Langerhans or a fraction thereof whereby insulin flows from the microcapsules, and said biocompatible semi-permeable membrane remains effective for a period of at least three months on implantation of said microcapsule to control blood sugar levels in the body in which the microcapsule is implanted.

5. The microcapsule of claim 4, wherein said islets of Langerhans are suspended in an aqueous medium.

9. The method of claim 4 wherein said reversibly-gellable water-soluble substance is a polysaccharide gum.

6. A method of encapsulating a core material within a semi-permeable membrane which is a hydrogel having an overall water content within the membrane structure of at least about 20 wt. %, which method 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 biocompatible semi-permeable membranes about the temporary capsules by contact between the temporary capsules and a biocompatible polyamino acid polymer containing free amino groups to cause ionic reaction with the acid groups in a surface layer of the capsule to provide a positively-charged surface, and

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

said semi-permeable membrane formation and said contact thereof with biocompatible polymeric material being such as to form microcapsules having a diameter of about 50 to about 2000 μ

m and a semi-permeable membrane thickness of about 5 to about 20 μ

m, and being such as to produce microcapsules capable of resisting degradation and remaining permeable in vivo for at least two months.

7. The method of claim 6 wherein said core material comprises living tissue which is in finely-divided suspended form in said aqueous solution in step (a).

8. The method of claim 7 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.

10. The method of claim 6 wherein said gum is an alkali metal alginate.

11. The method of claim 6 wherein said polymer-containing free amino groups has a molecular weight of about 11,000 to about 400,000 daltons.

12. The method of claim 6 wherein said polymer containing free amino groups comprises polylysine having a molecular weight of about 11,000 to about 100,000.

13. The method of claim 6 wherein said biocompatible polymeric material comprises a polysaccharide gum containing free acid groups.

14. The method of claim 6 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.

15. The method of claim 6 wherein said reversibly-gellable water-soluble substance comprises sodium alginate, said polymer containing free amino groups comprises polylysine, and said biocompatible polymeric material comprises sodium alginate.

18. The method of claim 6 including the further step of reliquifying the gel within the semi-permeable membrane.

16. The method of claim 20 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.

17. The method of claim 16, wherein said aqueous sodium alginate solution has a viscosity of at least about 30 cps.