Cell culture-containing tubular capsule produced by co-extrusion

• US 5,283,187 A
• Filed: 01/08/1991
• Issued: 02/01/1994
• Est. Priority Date: 11/17/1987
• Status: Expired due to Term

• Current Assignee: Brown University, Neurotech S.A. (LivaNova Plc)
• Original Assignee: Brown University
• Inventors: Wahlberg, Lars, Aebischer, Patrick
• Primary Examiner(s): Naff, David M.
• Application Number:
  US07/638,759
• Time in Patent Office: 1,120 Days
• Field of Search: 435/179, 435/180, 435/182, 435/240.22, 264/4, 264/4.7, 425/382 R, 425/382.2
• US Class Current: 435/182
• 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 2035/126 Immunoprotecting barriers, e.g. jackets, diffusion chambers
• CPC Class Codes: A61K 2035/128 capsules, e.g. microcapsules
• CPC Class Codes: A61K 35/12 Materials from mammals; Compositions comprising non-specified tissues or cells; Compositions comprising non-embryonic stem cells; Genetically modified cells vaccines or medicinal preparations containing antigens or antibodies A61K39/00
• CPC Class Codes: A61K 9/0024 Solid, semi-solid or solidifying implants, which are implanted or injected in body tissue compositions for intravenous administration, normal injectable solutions or dispersions for, e.g. subcutaneous administration A61K9/0019; brain implants A61K9/0085; (coated) prostheses, catheters or stents A61L
• CPC Class Codes: A61K 9/0085 Brain, e.g. brain implants; Spinal cord
• CPC Class Codes: A61K 9/0092 Hollow drug-filled fibres, tubes of the core-shell type, coated fibres, coated rods, microtubules or nanotubes
• CPC Class Codes: A61K 9/4816 Wall or shell material
• CPC Class Codes: A61K 9/4833 Encapsulating processes; Filling of capsules mechanical aspects A61J3/07
• CPC Class Codes: A61M 2210/0687 Skull, cranium
• CPC Class Codes: A61M 2210/0693 Brain, cerebrum
• CPC Class Codes: A61M 2230/005 Parameter used as control input for the apparatus
• CPC Class Codes: A61M 31/002 Devices for releasing a drug at a continuous and controlled rate for a prolonged period of time artificial gland structures or devices A61F2/022; intra-uterine contraceptive devices A61F6/14; tampons for introducing into the vagina A61F13/20, A61L15/00; suppositories or bougies for intra-vaginal or intra-uterine application A61K9/02; physical forms of medicinal preparations for sustained or differential drug release A61K9/20, A61K9/50
• CPC Class Codes: B01J 13/02 Making microcapsules or microballoons for medical preparations A61K9/50
• CPC Class Codes: B01J 13/04 by physical processes, e.g. drying, spraying
• CPC Class Codes: C12N 11/04 entrapped within the carrier, e.g. gel or hollow fibres
• CPC Class Codes: C12N 11/087 Acrylic polymers
• CPC Class Codes: C12N 11/098 formed in the presence of the enzymes or microbial cells
• CPC Class Codes: C12N 2533/30 Synthetic polymers
• CPC Class Codes: C12N 5/0012 Cell encapsulation
• CPC Class Codes: C12N 5/0614 : Adrenal gland
• CPC Class Codes: C12N 5/0619 : Neurons

Abstract

Living cells such as animal cells which produce biologically active factors are encapsulated within a semipermeable, polymeric membrane such as polyacrylate by co-extruding an aqueous cell suspension and a polymeric solution through a common port having at least one concentric bores to form a tubular extrudate having a polymeric membrane which encapsulates the cell suspension. The cell suspension is extruded through an inner bore and the polymeric solution is extruded through an outer bore while a pressure differential is maintained between the cell suspension and the polymeric solution to impede solvent diffusion from the polymeric solution into the cell suspension. The polymeric solution coagulates to form an outer coating or membrane as the polymeric solution and the cell suspension are extruded through the extrusion port. As the outer membrane is formed, the ends of the tubular extrudate are sealed to form a cell capsule. In one embodiment, the tubular extrudate is sealed at intervals to define separate cell compartments connected by polymeric links. In another embodiment, a cell capsule connected to a tethering filament is formed. The polymeric membrane may contain additives such as a surfactant, an anti-inflammatory agent or an anti-oxidant and can be coated with a protective barrier. The cell suspension may contain nutrients and an anchorage substrate.

Claims

1. A cell culturing capsule comprising a tubular, semipermeable, polymeric membrane encasing a viable cell culture, said tubular membrane being formed by

(i) co-extruding a viable cell culture and a polymeric solution through a common extrusion port having at least two concentric bores to form a tubular extrudate having a polymer outer coating which encases the cell culture, wherein the cell culture is extruded through an inner bore and the polymeric solution is extruded through an outer bore and a pressure differential is maintained between the cell culture and the polymeric solution during co-extrusion to impede solvent diffusion from the polymeric solution into the cell culture, and the polymeric solution and the cell culture are chosen so that coagulation of the polymeric solution occurs as the polymeric solution and cell culture are extruded through the extrusion port;

and

(ii) sealing the tubular extrudate to form at least one isolated tubular cell compartment.

2. The capsule of claim 1 wherein the polymeric membrane is permeable to molecules having a molecular weight of about 150,000 or less.

3. The capsule of claim 1 wherein the maximum outer diameter of the cell culture capsule ranges from about 0.1 to about 1.0 millimeters.

4. The capsule of claim 1 wherein the polymeric membrane has a membrane wall thickness ranging from about 10 to about 100 microns.

5. The capsule of claim 1 wherein the polymeric membrane further comprises a polyacrylate material.

6. The capsule of claim 1 wherein the polymeric membrane further comprises a surfactant.

7. The capsule of claim 1 wherein the polymeric membrane further comprises an anti-inflammatory agent.

8. The capsule of claim 1 wherein the polymeric membrane further comprises a anti-oxidant.

9. The capsule of claim 1 wherein the polymeric membrane further comprises a water-miscible solvent.

10. The capsule of claim 1 wherein the extruded tubular membrane is sealed at intervals to define separate cell compartments connected by polymeric links.

11. The capsule of claim 1 wherein the extruded tubular membrane is sealed to define at least one cell compartment connected to a tethering filament.

12. The capsule of claim 1 wherein the cell culture further comprises an aqueous cell suspension containing cells that secrete a biologically-active factor.

13. The capsule of claim 12 wherein the aqueous cell suspension further comprises a nutrient medium.

14. The capsule of claim 12 wherein the aqueous cell suspension further comprises an anchorage substrate material.

15. The capsule of claim 14 wherein the anchorage material comprises a collagen material.

16. The capsule of claim 14 wherein the anchorage material comprises laminin material.

17. The capsule of claim 14 wherein the anchorage material comprises a polyamino acid.

18. The capsule of claim 1 wherein capsule further comprises a protective barrier material which coats at least a portion of the outer surface of the polymeric membrane.

19. The capsule of claim 18 wherein the protective material is an inhibitor of protein interactions.

20. The capsule of claim 18 wherein the protective material is selected from the group consisting of polyethylene oxides, polypropylene oxide, derivatives and mixtures thereof.