Encapsulation of biological material

• US 4,352,883 A
• Filed: 03/28/1979
• Issued: 10/05/1982
• Est. Priority Date: 03/28/1979
• Status: Expired due to Term

• Current Assignee: Repligen Corporation
• Original Assignee: Damon Corporation (Thor Industries Incorporated)
• Inventors: Lim, Franklin
• Primary Examiner(s): Naff, David M.
• Application Number:
  US06/024,600
• Time in Patent Office: 1,287 Days
• Field of Search: 435/1, 435/182, 435/188, 435/262, 435/174, 435/175, 435/177, 435/178, 435/240, 424/31, 424/32, 424/34, 424/94, 424/101, 424/106, 424/110, 424/178, 264/4, 252/316, 3/1
• US Class Current: 435/178
• CPC Class Codes: A01N 1/02 Preservation of living parts
• CPC Class Codes: A01N 1/0231 Chemically defined matrices, e.g. alginate gels, for immobilising, holding or storing cells, tissue or organs for preservation purposes; Chemically altering or fixing cells, tissue or organs, e.g. by cross-linking, for preservation purposes
• 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 2035/128 capsules, e.g. microcapsules
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 38/00 Medicinal preparations containing peptides peptides containing beta-lactam rings A61K31/00; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, A61K31/00; ergot alkaloids of the cyclic peptide type A61K31/48; containing macromolecular compounds having statistically distributed amino acid units A61K31/74; medicinal preparations containing antigens or antibodies A61K39/00; medicinal preparations characterised by the non-active ingredients, e.g. peptides as drug carriers, A61K47/00
• CPC Class Codes: A61K 39/44 Antibodies bound to carriers
• 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: A61L 27/20 Polysaccharides
• CPC Class Codes: A61L 27/36 containing ingredients of undetermined constitution or reaction products thereof , e.g. transplant tissue, natural bone, extracellular matrix isolated natural compounds, e.g. collagen A61L27/24
• CPC Class Codes: B01J 13/04 by physical processes, e.g. drying, spraying
• CPC Class Codes: B01J 13/08 Simple coacervation, i.e. addition of highly hydrophilic material combined with spraying B01J13/043; combined with mechanical division B01J13/04
• CPC Class Codes: B01J 13/14 Polymerisation; cross-linking
• CPC Class Codes: B01J 13/16 Interfacial polymerisation
• CPC Class Codes: C12M 25/16 Particles; Beads; Granular material; Encapsulation chemical or physical processes conducted in the presence of fluids and solid particles B01J8/00
• CPC Class Codes: C12N 11/04 entrapped within the carrier, e.g. gel or hollow fibres
• CPC Class Codes: C12N 2533/30 Synthetic polymers
• CPC Class Codes: C12N 2533/32 : Polylysine, polyornithine
• CPC Class Codes: C12N 2533/74 : Alginate
• CPC Class Codes: C12N 5/0012 : Cell encapsulation
• CPC Class Codes: C12N 5/0671 : Three-dimensional culture, tissue culture or organ culture; Encapsulated cells
• CPC Class Codes: C12N 5/0677 : Three-dimensional culture, tissue culture or organ culture; Encapsulated cells

Abstract

A core material such as living tissue, individual cells, hormones, enzymes or antibodies is encapsulated in a semipermeable membrane that is permeable to small molecules for contact with the core material but is impermeable to potentially deleterious large molecules. Encapsulation may be carried out by suspending the core material in an aqueous medium containing a water soluble gum that can be reversibly gelled, forming the suspension into droplets, contacting the droplets with a solution of multivalent cations to gel the droplets as discrete, shape-retaining, water insoluble temporary capsules and cross-linking a surface layer of the temporary capsules to produce a semipermeable membrane around the capsules. Optionally the gel within the membrane may be reliquified by removing multivalent cations from the gel.

Claims

1. A process for encapsulating viable tissue within a semipermeable membrane, said process comprising the steps of:

A. suspending finely divided living tissue in an aqueous medium which is physiologically compatible with the tissue and which contains a water soluble substance which

(a) is physiologically compatible with the tissue;

and

(b) can be reversibly gelled to form a coherent, shape-retaining mass;

B. forming the suspension into droplets of a size sufficient to envelop tissue;

C. gelling the droplets to form discrete, shape-retaining temporary capsules;

D. forming a permanent semipermeable membrane about the temporary capsules; and

E. reliquifying the gel within the membrane.

2. A process for encapsulating a core material within a semipermeable membrane, said process comprising the steps of:

A. placing the material in a solution of a water-soluble substance that can be reversibly gelled;

B. forming the solution into droplets;

C. gelling the droplets to produce discrete shape-retaining temporary capsules;

D. forming semipermeable membranes about the temporary capsules; and

E. reliquifying the gel within said membranes.

3. The process of claim 2 wherein said substance comprises a gum.

4. The process of claim 3 wherein said gum has free acid groups and said membrane formation step is effected by contacting the temporary capsules with a polymer of a molecular weight between 3000 and 100,000 daltons and having free amino groups, said contacting being effective to form permanent polymer crosslinks between acid groups in a surface layer of the capsule.

5. The process of claim 4 wherein the polymer used for crosslinking is selected from the group consisting of polylysine and polyethylenimine, said polymer having an average molecular weight of about 35,000 daltons.

6. The process of claim 2 wherein the membrane is formed by an interfacial polymerization wherein the temporary capsules are used as a core material in the aqueous phase of a water-in-oil emulsion.

7. The process of claim 6 wherein the reactants used in the polymerizatiion are selected from the group consisting of water soluble polyols, diols, polyamines, and diamines, and water immiscible diacid halides, diacids, and multifunctional sulfonyl halides.

8. The process of claim 6 wherein the interfacial polymerization is a polyaddition reaction.

9. The process of claim 2 wherein the material is selected from the group consisting of enzymes, immunoproteins, activated charcoal particles, and viable tissue.

10. The process of claim 3 wherein the gum is an alkali metal alginate.

11. The process of claim 3 wherein the gum comprises a polysaccharide containing free acid groups.

12. The process of claim 2 wherein said core material is selected from the group consisting of hormones, enzymes and antibodies.

13. A process for encapsulating a core material within a membrane permeable to proteins of a molecular weight no greater than about 100 thousand daltons, said process comprising the steps of:

A. suspending the core material in an aqueous medium which contains a water-soluble gum containing acid groups;

B. forming the suspension into droplets;

C. subjecting the droplets to a solution of multivalent, cations to gel the droplets as discrete, shape-retaining, water insoluble temporary capsules; and

D. permanently cross-linking surface layers of said temporary capsules to produce a semipermeable membrane about said droplets by subjecting them to a polymer containing substitutents reactive with the acid groups of said gum, said polymer having a molecular weight within the range of 3000-100,000 daltons.

14. The process of claim 13 comprising the additional step of resolubilizing the gel within said capsules.

15. The process of claim 13 wherein the water soluble gum is sodium alginate and multivalent cation solution is a calcium solution.

16. The process of claim 15 including the additional step of removing the calcium ions contained within said capsules to resolubilize the gelled alginate interior of the membranes of said capsules.

17. The process of claim 15 wherein said core material is a mammalian tissue selected from the group consisting of Islets of Langerhans, liver, and individual cells thereof and said medium is a physiologically compatible tissue medium.

18. The process of claim 13 wherein said core material is a mammalian tissue selected from the group consisting of Islets of Langerhans, liver, and individual cells thereof and said medium is a physiologically compatible tissue medium.

19. The process of claim 13 wherein said core material comprises living tissue and said aqueous medium is a complete tissue culture medium sufficient to maintain said tissue in vitro.

20. The process of claim 13 wherein said cross-linking polymer is selected from the group consisting of polylysine and and polyethylenimine.

21. The process of claim 20 wherein the average molecular weight of said polymer is about 35,000 daltons.

22. The process of claim 13 wherein said forming step is effected by forcing droplets of the suspension through a capillary tube while vibrating the tube within the center of a vortex formed by imparting circular fluid motion to an aqueous solution containing multi-valent cations.