Matrix, cell implantation and method for their production and use

• US 20070166343A1
• Filed: 06/07/2004
• Published: 07/19/2007
• Est. Priority Date: 06/06/2003
• Status: Active Grant

• Current Assignee: Humanautocell GmbH
• Original Assignee: Humanautocell GmbH
• Inventors: Goerne, Martin, Kaufmann, Peter
• Granted Patent: US 7,618,646 B2
• Time in Patent Office: Days
• US Class Current: 424/423
• CPC Class Codes: A61L 27/26 Mixtures of macromolecular compounds
• CPC Class Codes: A61L 27/3804 characterised by specific cells or progenitors thereof, e.g. fibroblasts, connective tissue cells, kidney cells
• CPC Class Codes: A61L 27/3886 comprising two or more cell types
• CPC Class Codes: A61L 27/3895 using specific culture conditions, e.g. stimulating differentiation of stem cells, pulsatile flow conditions
• CPC Class Codes: A61L 27/56 Porous materials, e.g. foams or sponges
• CPC Class Codes: A61L 27/58 Materials at least partially resorbable by the body
• CPC Class Codes: A61P 1/16 for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
• CPC Class Codes: A61P 1/18 for pancreatic disorders, e.g. pancreatic enzymes
• CPC Class Codes: A61P 3/10 for hyperglycaemia, e.g. antidiabetics
• CPC Class Codes: A61P 35/00 Antineoplastic agents

Abstract

The invention relates to porous matrices based on a biologically compatible polymer or polymer mixture, to a cell implantation that is established on said matrices and to additional cell implantation based on cell mixtures of hepatocytes and islets of Langerhans. The invention also relates to a method for producing porous matrices, to matrices obtained according to said method and to a special method for obtaining cells for the inoculation of an implantable matrix.

Claims

1-45. -45. (canceled)

46. A porous matrix based on a biologically tolerated polymer or polymer mixture, wherein the matrix possesses pores having a size of 150 μ

m or less and pores having a size of 300 μ

m or more and the degree of porosity is from 93 to 98%.

47. The porous matrix as claimed in claim 46, wherein the matrix possesses pores having a size of 130 μ

m or less and pores having a size of 370 μ

m or more.

48. The porous matrix as claimed in claim 46, wherein about 0.5% to 6% of the pores have a mean diameter in the range of from 70 to 100 μ

m;

about 2% to 8% of the pores have a mean diameter in the range of from 101 to 115 μ

m;

about 2% to 8% of the pores have a mean diameter in the range of from 116 to 130 μ

m;

about 1% to 7% of the pores have a mean diameter in the range of from 131 to 300 μ

m;

about 11% to 23% of the pores have a mean diameter in the range of from 301 to 330 μ

m;

about 4% to 10% of the pores have a mean diameter in the range of from 331 to 360 μ

m;

about 5% to 17% of the pores have a mean diameter in the range of from 361 to 390 μ

m;

about 7% to 19% of the pores have a mean diameter in the range of from 391 to 420 μ

m;

about 3% to 9% of the pores have a mean diameter in the range of from 421 to 450 μ

m;

about 12% to 24% of the pores have a mean diameter in the range of from 451 to 480 μ

m; and

about 5% to 17% of the pores have a mean diameter in the range of from 481 to 510 μ

m.

49. The porous matrix as claimed in claim 46, wherein the biologically tolerated polymer is a biologically degradable polymer selected from the group consisting of natural polymers, albumin, fibrinogen, collagen, gelatin, chitin, chitosan, agarose, alginate, synthetic polymers, polyanhydrides, poly(ε

-caprolactone), poly(α

-hydroxyesters) and mixtures thereof.

50. The porous matrix as claimed in claim 49, wherein the biologically degradable polymer is poly(glycolic acid-lactic acid) having a lactic acid content of about 85 mol % and a glycolic acid content of about 15 mol %.

51. The porous matrix as claimed in claim 46, wherein the surface of the matrix is coated with at least one extracellular matrix protein selected from the group consisting of collagens, laminin and fibronectin.

52. The porous matrix as claimed in claim 51, wherein the coating contains fibronectin or the coating contains a mixture of type I collagen, laminin and type IV collagen.

74. An implant comprising the porous matrix as claimed in claim 46 and at least one cell.

75. The implant as claimed in claim 74 comprising a matrix based on a biologically tolerated polymer and cells of at least two cell types, wherein the cells of the first cell type are hepatocytes and the cells of the second cell type are islet of Langerhans cells.

76. The implant as claimed in claim 75, wherein the ratio of hepatocytes to islet of Langerhans cells is about 10⁶:

3000.

77. The implant as claimed in claim 75, wherein the ratio of hepatocytes to islet of Langerhans cells is about 10⁶:

3-200.

53. A method for preparing a porous matrix based on a biologically degradable polymer or polymer mixture, wherein a mixture comprising polymer particles having a grain size in the range of from about 20 to 950 μ

m and sodium chloride particles having a grain size in the range of from about 90 to 670 μ

m, is compacted and the sodium chloride is then dissolved from the mixture.

54. The method as claimed in claim 53, wherein the sodium chloride particle mixture comprises from 15 to 50% by weight of particles having a grain size of from 250 μ

m to 320 μ

m, from 20 to 65% by weight of particles having a grain size of from 330 to 380 μ

m, and from 15 to 62% by weight of particles having a grain size of from 390 μ

m to 425 μ

m.

55. The method as claimed in claim 53, wherein the sodium chloride particle mixture comprises from 1 to 15% by weight of particles having a grain size of from 108 μ

m to 140 μ

m, from 1 to 11% by weight of particles having a grain size of from 145 μ

m to 180 μ

m, from 3 to 21% by weight of particles having a grain size of from 185 μ

m to 220 μ

m, from 1 to 11% by weight of particles having a grain size of from 225 μ

m to 250 μ

m, from 15 to 50% by weight of particles having a grain size of from 250 μ

m to 320 μ

m, from 15 to 50% by weight of particles having a grain size of from 330 μ

m to 380 μ

m and from 5 to 29% by weight of particles having a grain size of from 390 μ

m to 425 μ

m.

56. The method as claimed in claim 53 wherein the polymer particle mixture comprises from 5 to 50% by weight of particles having a grain size of from 108 μ

m to 140 μ

m, from 10 to 55% by weight of particles having a grain size of from 145 μ

m to 180 μ

m, from 18 to 88% by weight of particles having a grain size of from 185 μ

m to 220 μ

m and from 5 to 45% by weight of particles having a grain size of from 225 μ

m to 250 μ

m.

57. The method as claimed in claim 53, wherein the ratio by weight of polymer particles to sodium chloride particles is from 1:

100 to 1;

10.

58. The method as claimed in claim 53, wherein a polymer solution is added to the mixture composed of polymer particles and sodium chloride particles, and the solvent is removed, prior to the compacting.

59. The method as claimed in claim 58, wherein the solvent dissolves the polymer but not the salt.

60. The method as claimed in claim 58, wherein the solvent is selected from the group consisting of acetone, ethyl acetate, methylene chloride, chloroform, hexafluoroisopropanol, chlorinated and fluorinated, aliphatic and aromatic hydrocarbons, tetrahydrofuran, ethyl methyl ketone, diethyl ketone and mixtures thereof.

61. The method as claimed in claim 58, wherein the polymer is poly(glycolic acid), poly(lactic acid) or poly(glycolic acid-lactic acid) and the solvent is chloroform.

62. The method as claimed in claim 58, wherein the ratio by weight of polymer particles to dissolved polymer is from 10:

1 to 1;

100.

63. The method as claimed in claim 53, wherein the compacting is effected by the action of pressure.

64. The method as claimed in claim 53, wherein water is allowed to act on the compacted mixture for the purpose of dissolving out the sodium chloride.

65. The method as claimed in claim 64, wherein the water is removed.

66. The method as claimed in claim 53, wherein the compacted mixture is first of all stored in a CO₂ atmosphere and the sodium chloride is subsequently dissolved from the compacted mixture.

73. A matrix obtained by the method as claimed in claim 53.

67. A method for preparing a porous matrix based on a biologically tolerated polymer or polymer mixture, wherein a mixture composed of polymer particles, particles of a porogenic material and a polymer solution is compacted and the porogenic material is then dissolved from the mixture.

68. The method as claimed in claim 67, wherein the polymer is at least one polymer selected from the group consisting of polyanhydrides, poly(orthoesters), poly(α

-hydroxyesters), poly(ester amides), polyamides, poly(ester ethers), poly-carbonates, polyalkylenes, polyalkylene glycols, polyalkylene oxides, polyalkylene terephthalates, polyvinyl alcohols, polyvinyl ethers, polyvinyl esters, polyvinyl halides, polyvinylpyrrolidones, polysiloxanes, polystyrenes, polyurethanes, derivatized celluloses and (meth)acrylic acid polymers and copolymers.

69. The method as claimed in claim 67, wherein the porogenic material is selected from water-soluble salts.

70. The method as claimed in claim 69, wherein the water-soluble salt is selected from the group consisting of sodium chloride, potassium chloride, sodium fluoride, potassium fluoride, sodium iodide, potassium iodide, sodium nitrate, sodium sulfate, sodium citrate, sodium tartrate, sugars and mixtures thereof.

71. The method as claimed in claim 67, wherein the solution contains polymer in dissolved form and polymer particles in solid form.

72. The method as claimed in claim 67, wherein the solution does not dissolve the porogenic material.