Human Therapeutic Cells Secreting Nerve Growth Factor
First Claim
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1. A human cell line comprising a stably integrated expression construct comprising the following sequences:
- a mammalian promoter,a coding sequence operably linked to said promoter, wherein said coding sequence codes for NGF, andan intron located in the transcript.
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Abstract
The present invention relates to human cell lines genetically modified to overexpress bioactive NGF. In another aspect the present invention relates to encapsulated human cell lines genetically modified to overexpress bioactive NGF, which can be used in therapy of Alzheimer'"'"'s disease, peripheral neuropathy and other neurological disorders amenable to local and prolonged NGF therapy.
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Citations
104 Claims
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1. A human cell line comprising a stably integrated expression construct comprising the following sequences:
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a mammalian promoter, a coding sequence operably linked to said promoter, wherein said coding sequence codes for NGF, and an intron located in the transcript. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33, 34, 35, 36, 37, 38, 39, 40, 41, 42, 43, 44, 45, 46, 47, 48, 49, 50, 51, 52, 53, 54, 55, 56, 57, 58, 59, 60, 61, 62, 63, 64, 65, 66, 67, 68, 69, 70, 71, 72, 73, 74, 75, 76, 77, 82, 83, 84, 85, 86, 87, 88, 89, 90, 92, 93, 94, 95, 96, 97)
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18. (canceled)
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78. (canceled)
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79. (canceled)
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81. (canceled)
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98. A method of treatment of Alzheimer'"'"'s Disease, comprising administering to an individual in need thereof a composition of ARPE-19 cells transfected with a vector selected from the group consisting of pCIn.hNGF, pCInKhNGF, and pNS1n.rINSintrA-hNGF, said composition being capable of secreting a therapeutically effective amount of mature human NGF.
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99. A method of treatment of Alzheimer'"'"'s Disease, comprising administering to an individual in need thereof a device, said device comprising a semipermeable membrane permitting the diffusion of human mature NGF therethrough, the device further comprising a composition of ARPE-19 cells transfected with a vector selected from the group consisting of pCIn.hNGF, pCInKhNGF, and pNS1n.rINSintrA-hNGF, said device being capable of releasing a therapeutically effective amount of mature human NGF.
- 100. An ARPE-19 cell line transfected with a vector selected from the group consisting of pCIn.hNGF, pCInKhNGF, and pNS1n.rINSintrA-hNGF.
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103. A composition of ARPE-19 cells being capable of secreting in excess of 10 ng of human mature NGF/ml/24 hours, when grown in confluent cultures in serumfree medium.
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104. An implantable cell device, the device comprising:
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(a) a semipermable membrane permitting the diffusion of human mature NGF therethrough; and (b) a population of ARPE-19 cells disposed within the semipermeable membrane, said device being capable of releasing in excess of 0.1 ng human mature NGF per 24 hours when grown in serumfree medium.
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Specification