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US5411883A Expired / Lapsed US ⏱️ 0.0 years remaining

Proliferated neuron progenitor cell product and process

Assignee: Somatix Therapy Corp
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Filing: 1992-08-12
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Granted: 1995-05-02
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Expires: 2012-08-12
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Abstract

Abstract This invention is based on the development of procedures for isolation and proliferation of neuron progenitor cells and is directed to growth, storage, production and implantation of proliferated neuron progenitor cells. The isolation and culture methods are designed to proliferate mammalian ventral mesencephalon neuron progenitor cells in vitro to produce a culture which differentiates to produce dopamine-producing cells. The products of this invention include a culture containing neuron progenitor cells, preferably, grown as aggregates in suspension cultures. The process of this invention for preparing neuron progenitor cells comprises obtaining ventral mesencephalon tissue from a donor at the appropriate stage of embryonic development; dissociation of the tissue to obtain single cells and small cell clusters for culture; culturing the neuron progenitor cells in an initial culture medium which selects for a novel cell culture containing neuron progenitor cells and growing the cells for a period of time in a second medium, during which the neuron progenitor cells proliferate.

Inventors

  • 👤 Barbara D. Boss
  • 👤 Dennis H. Spector

Bibliographic Details

Application Number —
Jurisdiction US
Data Source google_patents
Last Synchronized 2026-10-23 16:29
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/0618; C12N5/0623; A; A61; A61K; A61K35/00; A61K35/12; A61K35/30; C12N2501/00; C12N2501/01

Claims Analysis

Total Claims: 18
Claim 1
Independent Claim 99 words
A culture consisting essentially of mammalian ventral mesencephalon neuron progenitor cells or mammalian ventral mesencephalon neuron progenitor cells and their differentiated counterparts, said neuron progenitor cells being taken from a region of the brain that produces dopaminergic cells at a stage of development when the region of the brain has no dopamine-containing cells, said neuron progenitor cells in said culture having undergone at least one round of cell division after dissociation of tissue used to establish said culture, said progenitor cells in said culture being capable of seven to eight rounds of cell division after dissociation of said tissue.
Claim 2
Dependent Claim 99 words
A culture consisting essentially of mammalian ventral mesencephalon neuron progenitor cells or mammalian ventral mesencephalon neuron progenitor cells and their differentiated counterparts, said neuron progenitor cells being taken from a region of the brain that produces dopaminergic cells at a stage of development when the region of the brain has no dopamine-containing cells, said neuron progenitor cells in said culture having undergone at least one round of cell division after dissociation of tissue used to establish said culture, said progenitor cells in said culture being capable of seven to eight rounds of cell division after dissociation of said tissue.
Claim 3
Dependent Claim 11 words
The culture of claim 1 wherein said progenitor cells are porcine.
Claim 4
Dependent Claim 11 words
The culture of claim 1 wherein said progenitor cells are human.
Claim 5
Dependent Claim 19 words
The culture of claim 1 wherein said culture is capable of curing Parkinsonian symptoms about 4 months following implantation.
Claim 6
Dependent Claim 11 words
The culture of claim 1 wherein said culture comprises cell aggregates.
Claim 7
Dependent Claim 18 words
The culture of claim 5 wherein said aggregates range in size from about 100 to about 1000 microns.
Claim 8
Dependent Claim 17 words
The culture of claim 5 wherein said aggregates have loci of undifferentiated cells and loci of neurons.
Claim 9
Dependent Claim 34 words
A method for evaluating an agent which inhibits or enhances neuroblast proliferation or differentiation or for evaluating the toxicity of an agent on neurons comprising combining said agent with a culture of claim 1.
Claim 10
Independent Claim 112 words
A method for producing a culture comprising neuron progenitor cells comprising: a. obtaining ventral mesencephalon tissue from a mammalian embryo at Carnegie stages from 15 through 18; b. dissociation of said tissue to obtain single cells and small cell clusters for culture; c. culturing said single cells and small cell clusters in an initial culture medium which promotes the survival of neuron progenitor cells which are capable of proliferating in serum-free, defined medium for a sufficient period of time to produce a cell culture containing neuron progenitor cells capable of proliferating in serum-free, defined medium; and d. growing said cell culture containing neuron progenitor cells in a medium which maintains neuronal cells.
Claim 11
Independent Claim 112 words
A method for producing a culture comprising neuron progenitor cells comprising: a. obtaining ventral mesencephalon tissue from a mammalian embryo at Carnegie stages from 15 through 18; b. dissociation of said tissue to obtain single cells and small cell clusters for culture; c. culturing said single cells and small cell clusters in an initial culture medium which promotes the survival of neuron progenitor cells which are capable of proliferating in serum-free, defined medium for a sufficient period of time to produce a cell culture containing neuron progenitor cells capable of proliferating in serum-free, defined medium; and d. growing said cell culture containing neuron progenitor cells in a medium which maintains neuronal cells.
Claim 12
Dependent Claim 20 words
The method of claim 9 wherein said medium of step (c) is Ham's F12 medium supplemented with Chang's supplement C.
Claim 13
Dependent Claim 16 words
The method of claim 10 wherein said medium is additionally supplemented with glutamine and superoxide dismutase.
Claim 14
Dependent Claim 14 words
The method of claim 9 wherein said medium of step (d) is HN2 medium.
Claim 15
Dependent Claim 14 words
The method of claim 16 wherein said medium of step (d) is HN2 medium.
Claim 16
Dependent Claim 13 words
The method of claim 9 wherein said period of time is 5 days.
Claim 17
Dependent Claim 12 words
The method of claim 9 wherein said culture is an aggregate culture.
Claim 18
Dependent Claim 16 words
The method of claim 14 wherein not more than 10 5 cells are seeded per culture.

References Cited (Backward Citations)

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3.5 Years (Year 4) 1998-05-01 1998-10-31 1999-05-02 $2000.00 Window Closed / Lapsed
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