Amnion-derived cell compositions, methods of making and uses thereof

• US 20090075381A1
• Filed: 07/17/2008
• Published: 03/19/2009
• Est. Priority Date: 03/31/2005
• Status: Active Grant

• Current Assignee: Stemnion, Inc.
• Original Assignee: Stemnion, Inc.
• Inventors: Smith, Charlotte A., Marshall, Vivienne S., Clarke, Diana L., Banas, Richard A.
• Granted Patent: US 8,278,095 B2
• Time in Patent Office: Days
• US Class Current: 435/378
• CPC Class Codes: A61K 2800/10 General cosmetic use
• 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 35/50 Placenta; Placental stem cells; Amniotic fluid; Amnion; Amniotic stem cells
• CPC Class Codes: A61K 8/982 Reproductive organs; Embryos, Eggs
• CPC Class Codes: A61Q 19/00 Preparations for care of the skin
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2502/02 embryonic cells
• CPC Class Codes: C12N 5/0605 Cells from extra-embryonic tissues, e.g. placenta, amnion, yolk sac, Wharton's jelly
• CPC Class Codes: C12N 5/0629 Keratinocytes; Whole skin

Abstract

The invention is directed to substantially purified amnion-derived cell populations, compositions comprising the substantially purified amnion-derived cell populations, and to methods of creating such substantially purified amnion-derived cell populations, as well as methods of use. The invention is further directed to antibodies, in particular, monoclonal antibodies, that bind to amnion-derived cells or, alternatively, to one or more amnion-derived cell surface protein markers. The invention is further directed to methods for producing the antibodies, methods for using the antibodies, and kits comprising the antibodies.

Claims

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12. A method of obtaining a substantially purified population of amnion-derived cells, comprising:

a) providing a population of amnion-derived cells;

b) contacting the cells with

(i) one or more antibodies selected from the group consisting of anti-CD105, anti-CD90, anti-CD117, anti-CD140b, anti-CD34, anti-CD44, and anti-CD45, antibodies; and

(ii) one or more antibodies selected from the group consisting of anti-CD29, anti-CD9, anti-CD10, anti-CD26, anti-CD71, anti-CD166, anti-CD227, anti-EGF-R, anti-SSEA-4, and anti-HLA-G antibodies; and

c) separating the cells that do not bind to the antibodies of (i) from the cells that do bind to the antibody of (i) and separating the cells that do not bind to the antibodies of (ii) from the cells that do bind to the antibody of (ii) such that a substantially purified population of amnion-derived cells that do not bind to the antibodies of (i) and do bind to the antibody of (ii) is obtained

60. The method of claim 12 wherein the antibodies of (i) are anti-CD105, anti-CD90, anti-CD117, anti-CD140b, anti-CD34, anti-CD44, and anti-CD45 antibodies, and the antibodies of (ii) are anti-CD29, anti-CD9, anti-CD10, anti-CD26, anti-CD71, anti-CD166, anti-CD227, anti-EGF-R, anti-SSEA-4, and anti-HLA-G antibodies.

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16. A composition comprising conditioned medium obtained from a substantially purified population of amnion-derived cells that is negative for expression of the protein markers CD90 and CD117.

63. The composition of claim 16 wherein the substantially purified population of amnion-derived cells is further negative for expression of the protein marker CD105.

64. The composition of claim 16 wherein the substantially purified population of amnion-derived cells is positive for expression of the protein marker CD29.

65. The composition of claim 16 wherein the substantially purified population of amnion-derived cells is positive for expression of HLA-G.

66. The composition of claim 16 wherein the substantially purified population of amnion-derived cells is further negative for the expression of telomerase.

67. The composition of claim 66 wherein the substantially purified population of amnion-derived cells is positive for expression of HLA-G.

68. The composition of claim 16, which is a pharmaceutical composition.

69. The composition of claim 16, wherein the amnion-derived cells are an expanded amnion-derived cell composition.

70. The composition of claim 16, wherein the conditioned medium is diluted with unconditioned medium.

17. A composition comprising cell lysate obtained from a substantially purified population of amnion-derived cells that is negative for expression of the protein markers CD90 and CD117.

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59. A method of obtaining a substantially purified population of amnion-derived cells, comprising:

a) providing a population of amnion-derived cells;

b) contacting the cells with anti-CD90 and anti-CD117 antibodies; and

c) separating the cells that do not bind to the antibodies of (b) from the cells that do bind to the antibody of (b) such that a substantially purified population of amnion-derived cells that do not bind to the antibodies of (b) is obtained.

62. The method of claim 59 further optionally comprising:

d) contacting the cells of c) with anti-HLA-G antibodies; and

e) separating the cells that bind to the anti-HLA-G antibodies from the cells that do not bind to the anti-HLA-G antibodies such that a substantially purified population of amnion-derived cells that bind to the anti-HLA-G antibodies is obtained.

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71. A method of making a composition comprising conditioned media, comprising

a) seeding amnion-derived cells in culture flasks,

b) culturing the cells in animal-free medium supplemented with EGF,

c) culturing the cells until they reach near confluency,

d) removing the medium and applying fresh medium, and

e) collecting the freshly applied medium after culture.

72. The method of claim 71 wherein the medium does not contain serum.

73. The method of claim 72 wherein the medium is IMDM medium.

74. The method of claim 71 wherein the freshly applied medium of step (e) is collected after at least one day in culture.

75. A composition comprising conditioned media made by the method of claim 71.

76. A composition comprising conditioned media which is positive for expression of Angiopoietin-2, Angiogenin, bFGF, EGF, FGF-7, FGF-4, IGF-1, IL-1 beta, IL-2, IL-4, IL-6, IL-8, IL-10, PDGF-AA, PDGF-AB, PDGF-BB, PDGF-Ra, and PDGF-Rb as detected by antibody array.

77. The composition of claim 76 which is negative for the expression of TGF-a, TGF-beta 1, TGF-beta 2, and TGF-beta 3 as detected by antibody array.

78. A composition comprising conditioned media obtained from cultured amnion-derived cells.