Methods of use of uncoated gel particles

• US 5,912,005 A
• Filed: 11/19/1996
• Issued: 06/15/1999
• Est. Priority Date: 04/15/1994
• Status: Expired due to Fees

• Current Assignee: Biohybrid Technologies Incorporated
• Original Assignee: Biohybrid Technologies Incorporated
• Inventors: Lanza, Robert P., Kuhtreiber, Willem M., Chick, William L.
• Primary Examiner(s): CLARDY, S
• Application Number:
  US08/746,970
• Time in Patent Office: 938 Days
• Field of Search: 435/174, 435/177, 435/240.22, 435/240.43, 435/243, 435/382, 264/4.3, 424/422, 424/423, 424/424, 424/489, 514/866, 514/907, 514/885, 514/953
• US Class Current: 424/424
• CPC Class Codes: A61K 2035/128 capsules, e.g. microcapsules
• CPC Class Codes: A61K 35/15 Cells of the myeloid line, e.g. granulocytes, basophils, eosinophils, neutrophils, leucocytes, monocytes, macrophages or mast cells; Myeloid precursor cells; Antigen-presenting cells, e.g. dendritic cells presenting a specific antigen A61K39/00; therapeutic combinations of antibodies, or fragments thereof, and blood-derived cells A61K39/00
• CPC Class Codes: A61K 35/17 Lymphocytes; B-cells; T-cells; Natural killer cells; Interferon-activated or cytokine-activated lymphocytes when activated by a specific antigen A61K39/00
• CPC Class Codes: A61K 35/39 Pancreas; Islets of Langerhans Langerhans cells of epidermis A61K35/36
• CPC Class Codes: A61K 38/30 Insulin-like growth factors, i.e. somatomedins, e.g. IGF-1, IGF-2 insulin-like growth factor binding protein A61K38/1754
• CPC Class Codes: A61K 38/4846 Factor VII (3.4.21.21); Factor IX (3.4.21.22); Factor Xa (3.4.21.6); Factor XI (3.4.21.27); Factor XII (3.4.21.38)
• CPC Class Codes: A61K 48/00 Medicinal preparations containing genetic material which is inserted into cells of the living body to treat genetic diseases; Gene therapy
• 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: A61P 13/02 of urine or of the urinary tract, e.g. urine acidifiers
• CPC Class Codes: A61P 15/00 Drugs for genital or sexual disorders for disorders of sex hormones A61P5/24; Contraceptives
• CPC Class Codes: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/08 Antiepileptics; Anticonvulsants
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 3/02 Nutrients, e.g. vitamins, minerals
• CPC Class Codes: A61P 3/08 for glucose homeostasis pancreatic hormones A61P5/48
• CPC Class Codes: A61P 37/00 Drugs for immunological or allergic disorders
• CPC Class Codes: A61P 37/06 Immunosuppressants, e.g. drugs for graft rejection
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 7/00 Drugs for disorders of the blood or the extracellular fluid
• CPC Class Codes: C12N 11/04 entrapped within the carrier, e.g. gel or hollow fibres
• CPC Class Codes: C12N 2533/74 : Alginate
• CPC Class Codes: C12N 5/0012 : Cell encapsulation
• CPC Class Codes: C12N 5/0677 : Three-dimensional culture, tissue culture or organ culture; Encapsulated cells

Abstract

The invention covers a method of implanting a living donor cell into a host animal without inflammatory response or rejection of the donor cell by the host animal, by obtaining an uncoated particle of a biocompatible, temperature-independent gel that encapsulates the living donor cell, wherein the uncoated particle provides a molecular weight cutoff that prevents host animal immune cells from entering the particle, yet does not have to prevent entry of host animal IgG and complement into the particle, and implanting the uncoated particle into the host animal.

Claims

1. A method of implanting a living donor cell into a host animal, wherein said animal is a mammal larger than a rat or mouse, with minimal inflammatory response or rejection of the donor cell by the host animal, said method comprising the steps of

obtaining an uncoated particle comprising a biocompatible, temperature-independent gel that encapsulates the living donor cell, wherein said uncoated particle provides a molecular weight cutoff that prevents host animal immune cells from entering the particle, and does not prevent entry of host animal IgG and complement into said particle, wherein said particle has a diameter of 700-6,000 microns, and

implanting the uncoated particle into the host animal.

2. A method of claim 1, wherein said donor cell is obtained from an species that is the same as the host animal.

3. A method of claim 2, wherein the host animal is a dog.

4. A method of claim 2, wherein the host animal is a human.

5. A method of claim 4, wherein the donor cell is a genetically altered human cell.

6. A method of claim 1, wherein the donor cell is obtained from an animal species that is different from the host animal.

7. A method of claim 6, wherein the host animal is a dog.

8. A method of claim 6, wherein the host animal is a human.

9. A method of claim 8, wherein the donor cell is a porcine, bovine, or canine cell.

10. A method of claim 8, wherein the donor cell is a pancreatic islet cell.

11. A method of claim 1, wherein the donor cell secretes Factor IX, Factor VIII, an interleukin, an interferon, or an endocrine hormone.

12. A method of claim 1, wherein the donor cell secretes a nerve growth factor, tumor necrosis factor alpha, a neurotropic factor, or a neurotransmitter.

13. A method of claim 1, wherein said gel particle is spherical and has a diameter of from 1,500-3,500 microns.

14. A method of claim 13, wherein said gel particle has a diameter of from 2000 to 4500 microns.

15. A method of claim 1, wherein said gel is an alginate or alginate derivative.

16. A method of claim 15, wherein said alginate is crosslinked with an ion.

17. A method of claim 16, wherein said alginate is crosslinked with a calcium salt.

18. A method of claim 1, wherein said uncoated gel particle is biodegradable.

19. A method of claim 18, wherein a rate of degradation of said gel in said uncoated particle is selected to match a life expectancy of said donor cell.

20. A method of claim 1, wherein said uncoated particle encapsulates an autologous erythrocyte in addition to the donor cell.

21. A method of claim 1, wherein said uncoated particle containing a living cell is treated with a nitric oxide inhibitor prior to implantation.

61. A method of claim 1, wherein said particle is other than a sphere.

22. A method of claim l, further comprising the step of administering a drug to the host animal at a dosage effective to inhibit fibrosis and inflammation around said uncoated particle, but at a dosage lower than that required to achieve immunosuppression when said donor cell is implanted into the host animal without encapsulation.

23. A method of claim 22, wherein said drug is cyclosporin A and is administered at a dosage that achieves a whole blood trough level of less than about 100 ng/ml in the host animal.

24. A method of claim 22, wherein said drug is administered for up to one month after implantation, and is then no longer administered.

45. A method of claim 22, wherein said drug is a nonsteroidal anti-inflammatory drug.

25. A method of implanting a living donor cell into a host animal, wherein said animal is a mammal larger than a rat or mouse, with inflammatory response or rejection of the donor cell by the host animal, said method comprising the steps of

suspending the living donor cell in a liquid medium, said medium comprising water and a biocompatible, temperature-independent liquid gel,

forming a droplet of said liquid medium that contains at least one living cell,

solidifying said droplet to form a gel particle that encapsulates the living cell, whereby no outer coating is formed on said particle, and wherein said uncoated particle provides a molecular weight cutoff that prevents host animal immune cells from entering the particle, and does not prevent entry of host animal IgG and complement into said particle, wherein said particle has a diameter of 700-6,000 microns and

implanting the uncoated particle into the host.

26. A method of claim 25, wherein said liquid medium contains pancreatic islets.

27. A method of claim 25, wherein said pancreatic islets are contained in said liquid medium at a density of 10,000 to 35,000 islets per milliliter of said medium.

28. A method of claim 25, wherein said liquid medium contains living cells at a density of about 10⁵ to 10⁸ cells per milliliter of said medium.

62. A method of claim 25, wherein said particle is other than a sphere.

29. A method of treating a disease in a patient, wherein said patient is a mammal larger than a rat or mouse, caused by a deficient production of a substance in the patient, said method comprising the steps of

obtaining an uncoated particle comprising a biocompatible, temperature-independent gel that encapsulates a living donor cell that secretes said substance, wherein said uncoated particle provides a molecular weight cutoff that prevents patient immune cells from entering the particle, and does not prevent entry of patient IgG and complement into said particle, wherein said particle has a diameter of 700-6,000 microns, and

implanting the uncoated particle into the patient in a location and in a manner that allows the living cell to remain physiologically active and secrete said substance into the patient to treat said disease.

30. A method of claim 29, wherein the donor cell is obtained from an animal species that is the same as the patient.

31. A method of claim 30, wherein the patient is a dog.

32. A method of claim 30, wherein the patient is a human.

33. A method of claim 32, wherein the donor cell is a genetically altered human cell.

34. A method of claim 29, wherein the donor cell is obtained from a species that is different from the patient.

35. A method of claim 34, wherein the patient is a dog.

36. A method of claim 34, wherein the patient is a human.

37. A method of claim 29, wherein the donor cell is a porcine, bovine, canine, bacterial, fungal, or plant cell.

38. A method of claim 29, wherein the disease is diabetes and the donor cell is a pancreatic islet cell.

39. A method of claim 29, wherein the donor cell secretes Factor IX, Factor VIII, an interleukin, an interferon, an endocrine hormone, a nerve growth factor, tumor necrosis factor alpha, a neurotropic factor or a neurotransmitter.

40. A method of claim 29, wherein the disease is diabetes mellitus, hepatic disease, amyotrophic lateral sclerosis, hemophilia, hypothyroidism, Parkinson'"'"'s disease, acquired immune deficiency syndrome, Duchenne'"'"'s muscular dystrophy, infertility, epilepsy, Huntington'"'"'s disease, hypoparathyroidism, a mood disorder, a motor neuron disease, osteoporosis, or Alzheimer'"'"'s disease.

41. A method of claim 29, wherein the gel particles are implanted into an immunoprivileged site in the patient.

63. A method of claim 29, wherein said particle is other than a sphere.

42. An in vivo method of culturing a living cell, said method comprising the steps of

encapsulating the living cell in an uncoated particle comprising a biocompatible, temperature-independent gel,

inserting said uncoated particle into an animal, and

allowing said animal to thrive, thereby culturing the cell wherein said animal is a mammal larger than a rat or mouse, wherein said particle has a diameter of 700-6,000 microns and wherein said uncoated particle provides a molecular weight cutoff that prevents host animal immune cells from entering the particle, and does not prevent entry of host animal IgG and complement into said particle.

43. An in vitro method of culturing a living cell, said method comprising the steps of

encapsulating the living cell in an uncoated particle comprising a biocompatible, temperature-independent gel, wherein said particle has a diameter of 700-6,000 microns and wherein said uncoated particle provides a molecular weight cutoff that prevents host animal immune cells from entering the particle, and does not prevent entry of host animal IgG and complement into said particle

placing said uncoated particle into a medium including nutrients and oxygen, and

maintaining a sufficient amount of nutrients and oxygen in said medium to allow the cell to thrive, thereby culturing the cell.

44. A method of manufacturing uncoated, temperature-independent gel particles containing living cells consisting of the steps of

suspending the living cells in a liquid medium, said medium comprising water and a biocompatible, temperature-independent, liquid gel,

forming a droplet of said liquid medium,

solidifying said droplet to form a gel particle that encapsulates the living cells, whereby no outer coating is formed on said particle, wherein said particle has a diameter of 700-6,000 μ

m and wherein said uncoated particle provides a molecular weight cutoff that prevents host animal immune cells from entering the particle, and does not prevent entry of host animal IgG and complement into said particle, and

storing said gelled uncoated particles in a nutrient medium to maintain the viability of the living cells.

46. A method of implanting a living donor cell into a host animal, wherein said animal is a mammal larger than a rat or mouse, with minimal inflammatory response or rejection of the donor cell by the host animal, said method comprising the steps of

obtaining an uncoated particle comprising a biocompatible, temperature-independent gel that encapsulates the living donor cell, wherein said uncoated particle provides a molecular weight cutoff that prevents host animal immune cells from entering the particle, and does not prevent entry of host animal IgG and complement into said particle,

implanting the uncoated particle into the host animal, and

administering a drug to the host animal at a dosage effective to inhibit fibrosis and inflammation of the uncoated particle, but at a dosage lower than that required to achieve immunosuppression when the donor cell is implanted into the host animal without encapsulation.

47. A method of claim 46, wherein said donor cell is obtained from an species that is the same as the host animal.

48. A method of claim 46, wherein the host animal is a human.

49. A method of claim 46, wherein the donor cell is a genetically altered human cell.

50. A method of claim 46, wherein the donor cell is obtained from an animal species that is different from the host animal.

51. A method of claim 50, wherein the host animal is a human.

52. A method of claim 50, wherein the donor cell is a porcine, bovine, or canine cell.

53. A method of claim 46, wherein the donor cell is a pancreatic islet cell.

54. A method of claim 46, wherein the donor cell secretes Factor IX, Factor VIII, an interleukin, an interferon, or an endocrine hormone.

55. A method of claim 46, wherein the donor cell secretes a nerve growth factor, tumor necrosis factor alpha, a neurotropic factor, or a neurotransmitter.

56. A method of claim 46, wherein said gel particle is spherical and has a diameter of from 50 to 6000 microns.

57. A method of claim 46, wherein said gel is an alginate or alginate derivative.

58. A method of claim 46, wherein said drug is cyclosporin A and is administered at a dosage that achieves a whole blood trough level of less than about 100 ng/ml in the host animal.

59. A method of claim 46, wherein said drug is administered for up to one month after implantation, and is then no longer administered.

60. A method of claim 46, wherein said drug is a nonsteroidal antis inflammatory drug.

64. A method of claim 46, wherein said particle is other than a sphere.