US6833269B2
Expired / Lapsed
US
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Making neural cells for human therapy or drug screening from human embryonic stem cells
Assignee: Asterias Biotherapeutics Inc
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Filing: 2001-05-31
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Granted: 2004-12-21
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Expires: 2021-05-31
Abstract
Abstract This invention provides populations of neural progenitor cells, differentiated neurons, glial cells, and astrocytes. The populations are obtained by culturing stem cell populations (such as embryonic stem cells) in a cocktail of growth conditions that initiates differentiation, and establishes the neural progenitor population. The progenitors can be further differentiated in culture into a variety of different neural phenotypes, including dopaminergic neurons. The differentiated cell populations or the neural progenitors can be generated in large quantities for use in drug screening and the treatment of neurological disorders.
Inventors
- 👤 Melissa K. Carpenter
Bibliographic Details
Application Number
—
Jurisdiction
US
Data Source
google_patents
Last Synchronized
2026-11-02 23:26
CPC Classifications
C; C12; C12N; C12N5/00; C12N5/06; C12N5/0602; C12N5/0618; A; A61; A61P; A61P25/00; A61P25/08; A61P25/14; A61P25/16; A61P25/28; A61P43/00; A61P9/00; A61P9/08; A61P9/10; C12N5/0623; C12Q; C12Q1/00; C12Q1/68; C12Q1/6876; C12Q1/6881; A61K; A61K35/00; A61K35/12; C12N2501/00; C12N2501/01; C12N2501/10; C12N2501/105; C12N2501/11; C12N2501/115; C12N2501/119; C12N2501/13; C12N2501/135; C12N2503/00; C12N2503/02; C12N2506/00; C12N2506/02; C12N2510/00; C12Q2600/00; C12Q2600/158
Claims Analysis
Total Claims: 45
Claim 1
Independent Claim
52 words
A method for producing a neural cell population from human embryonic stem (hES) cells, comprising culturing hES cells in a medium containing one or more added neurotrophins and one or more added nitrogens, thereby producing a population in which at least 60% of the cells express A2B5, polyslalylated NCAM, MAP-2, or Nestin.
Claim 2
Independent Claim
52 words
A method for producing a neural cell population from human embryonic stem (hES) cells, comprising culturing hES cells in a medium containing one or more added neurotrophins and one or more added nitrogens, thereby producing a population in which at least 60% of the cells express A2B5, polyslalylated NCAM, MAP-2, or Nestin.
Claim 3
Dependent Claim
19 words
The method of claim 1 , wherein the added neurotrophins include neurotrophin 3 (NT-3) or brain-derived neurotrophic factor (BDNF).
Claim 4
Dependent Claim
15 words
The method of claim 2 , wherein the added neurotrophins include both NT-3 and BDNF.
Claim 5
Dependent Claim
34 words
The method at claim 1 , wherein the added mitogens include a mitogen selected from epidermal growth factor (EGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF), and insulin-like growth factor 1 (IGF-1).
Claim 6
Dependent Claim
15 words
The method of claim 4 , wherein the added mitogens include both PDGF and IGF-1.
Claim 7
Dependent Claim
14 words
The method of claim 1 , wherein the hES cells are cultured on fibronectin.
Claim 8
Dependent Claim
17 words
The method of claim 1 , comprising selecting and propagating cells that are positive for polyslalylated NCAM.
Claim 9
Dependent Claim
19 words
The method of claim 1 , wherein the produced cell population is at least 60% positive for polystalylated NCAM.
Claim 10
Dependent Claim
19 words
The method of claim 1 , wherein the produced cell population is at least 90% positive for polystalylated NCAM.
Claim 11
Dependent Claim
18 words
The method of claim 1 , wherein the produced cell population is at least 38% β-tubulin III positive.
Claim 12
Dependent Claim
38 words
The method of claim 1 , wherein the produced cell population has the characteristic that culturing it in the presence of neurotrophins maintains capacity of the cells to proliferate and to form tyrosine hydroxylase positive neurons upon differentiation.
Claim 13
Dependent Claim
18 words
The method of claim 1 , wherein the produced cell population is essentially free of undifferentiated hES cells.
Claim 14
Dependent Claim
20 words
The method of claim 1 , further comprising differentiating the cells into a population that contains at least 30% neurons.
Claim 15
Dependent Claim
22 words
The method of claim 1 , further comprising differentiating the cells into a population that contains at least 30% MAP-2 positive cells.
Claim 16
Dependent Claim
23 words
The method of claim 14 , wherein at least 3% of the differentiated cells positive for MAP-2 are also positive for tyrosine hydroxylase.
Claim 17
Dependent Claim
23 words
The method of claim 14 , wherein at least 8% of the differentiated cells positive for MAP-2 are also positive for tyrosine hydroxylase.
Claim 18
Dependent Claim
39 words
The method of claim 14 , wherein the differentiating comprises culturing the cells in a medium containing one or more factors selected from neurotrophins, cAMP, and ascorbic acid for at least 3 days, in the absence of added mitogens.
Claim 19
Dependent Claim
16 words
The method of claim 1 , wherein the differentiated cell population comprises sensory or motor neurons.
Claim 20
Dependent Claim
15 words
The method of claim 1 , wherein the differentiated cell population comprises oligodendrocytes or astrocytes.
Claim 21
Dependent Claim
18 words
The method of claim 1 , further comprising combining the produced cell population with a pharmaceutically compatible excipient.
Claim 22
Independent Claim
67 words
A method for producing a neural cell population from human embryonic stem (hES) cells, comprising initialing differentiation of the hES cells into a heterogeneous population of precursor cells, then culturing the initiated cells in a medium containing one or more added neurotrophins and one or more added mitogens, thereby producing a population in which at least 60% of the cells express A2B5, polysialylated NCAM, MAP-2, or Nestin.
Claim 23
Independent Claim
67 words
A method for producing a neural cell population from human embryonic stem (hES) cells, comprising initialing differentiation of the hES cells into a heterogeneous population of precursor cells, then culturing the initiated cells in a medium containing one or more added neurotrophins and one or more added mitogens, thereby producing a population in which at least 60% of the cells express A2B5, polysialylated NCAM, MAP-2, or Nestin.
Claim 24
Dependent Claim
19 words
The method of claim 21 , wherein the added neurotrophins include neurotrophin 3 (NT-3) or brain-derived neurotrophic factor (BDNF).
Claim 25
Dependent Claim
15 words
The method of claim 22 , wherein the added neurotrophins include both NT-3 and BDNF.
Claim 26
Dependent Claim
34 words
The method of claim 21 , wherein the added mitogens include a mitogen selected from epidermal growth factor, (EGF), basic fibroblast growth factor (bFGF), platelet-derived growth factor (PDGF), and insulin-like growth factor 1 (IGF-1).
Claim 27
Dependent Claim
15 words
The method of claim 24 , wherein the added mitogens include both PDGF and IGF-1.
Claim 28
Dependent Claim
14 words
The method of claim 21 , wherein the precursor cells are cultured on fibronectin.
Claim 29
Dependent Claim
28 words
The method of claim 21 , wherein differentiation is initiated by culturing the hES cells in a medium containing retinoic acid before culturing with the neurotrophins and mitogens.
Claim 30
Dependent Claim
24 words
The method of claim 21 , wherein differentiation is initiated by culturing the cells as embryold bodies before culturing with the neurotrophins and mitogens.
Claim 31
Dependent Claim
24 words
The method of claim 21 , wherein differentiation is initiated by culturing the cells as cell aggregates before culturing with the neurotrophins and mitogens.
Claim 32
Dependent Claim
17 words
The method of claim 21 , comprising selecting and propagating cells that are positive for polysialylated NCAM.
Claim 33
Dependent Claim
19 words
The method of claim 21 , wherein the produced cell population is at least 60% positive for polysialytated NCAM.
Claim 34
Dependent Claim
19 words
The method of claim 21 , wherein the produced cell population is at least 90% positive for polysialylated NCAM.
Claim 35
Dependent Claim
18 words
The method of claim 21 , wherein the produced cell population is at least 38% β-tubulin III positive.
Claim 36
Dependent Claim
38 words
The method of claim 21 , wherein the produced cell population has the characteristic that culturing it in the presence of neurotrophins maintains capacity of the cells to proliferate and to form tyrosine hydroxylase positive neurons upon differentiation.
Claim 37
Dependent Claim
18 words
The method of claim 21 , wherein the produced cell population is essentially free of undifferentiated hES cells.
Claim 38
Dependent Claim
20 words
The method of claim 21 , further comprising differentiating the cells into a population that contains at least 30% neurons.
Claim 39
Dependent Claim
22 words
The method of claim 21 , further comprising differentiating the cells into a population that contains at least 30% MAP-2 positive cells.
Claim 40
Dependent Claim
23 words
The method of claim 37 , wherein at least 3% of the differentiated cells positive for MAP-2 are also positive for tyrosine hydroxylase.
Claim 41
Dependent Claim
23 words
The method of claim 37 , wherein at least 8% of the differentiated cells positive for MAP-2 are also positive for tyrosine hydroxylase.
Claim 42
Dependent Claim
39 words
The method of claim 37 , wherein the differentiating comprises culturing the cell in a medium containing one or more factors selected from neurotrophins, cAMP, and ascorbic acid for at least 3 days, in the absence of added mitogens.
Claim 43
Dependent Claim
16 words
The method of claim 21 , wherein the differentiated cell population comprises sensory or motor neurons.
Claim 44
Dependent Claim
15 words
The method of claim 21 , wherein the differentiated cell population comprises oligodendrocytes or astrocytes.
Claim 45
Dependent Claim
18 words
The method of claim 21 , further comprising combining the produced cell population with a pharmaceutically compatible excipient.
References Cited (Backward Citations)
US5981165A
Cited
US5851832A
Cited
US5672499A
Cited
US5849553A
Cited
US5766948A
Cited
US5639618A
Cited
US5698829A
Cited
US6040180A
Cited
WO1998050526A1
Cited
WO1999001159A1
Cited
WO1999004775A2
Cited
US5968829A
Cited
WO1999028443A1
Cited
WO2000017323A1
Cited
WO2000047762A2
Cited
US6238922B1
Cited
WO2001068815A1
Cited
WO2001098463A1
Cited
USPTO Annuity & Maintenance Windows
| Annuity Stage | Window Opens | Payment Deadline | Surcharge Period Until | Estimated Fee | Status |
|---|---|---|---|---|---|
| 3.5 Years (Year 4) | 2007-12-21 | 2008-06-21 | 2008-12-21 | $2000.00 | Window Closed / Lapsed |
| 7.5 Years (Year 8) | 2011-12-21 | 2012-06-21 | 2012-12-21 | $3760.00 | Window Closed / Lapsed |
| 11.5 Years (Year 12) | 2015-12-21 | 2016-06-21 | 2016-12-21 | $7700.00 | Window Closed / Lapsed |