Neural progenitor cell populations

• US 8,148,148 B2
• Filed: 11/16/2005
• Issued: 04/03/2012
• Est. Priority Date: 05/17/2000
• Status: Expired due to Fees

• Current Assignee: Asterias Biotherapeutics Incorporated (Lineage Cell Therapeutics, Inc.)
• Original Assignee: Geron Corporation
• Inventors: Carpenter, Melissa K.
• Primary Examiner(s): FALK, ANNE MARIE
• Application Number:
  US11/281,040
• Publication Number:
  US 20060078545A1
• Time in Patent Office: 2,330 Days
• Field of Search: None
• US Class Current: 435/368
• 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: A61P 25/00 Drugs for disorders of the nervous system
• CPC Class Codes: A61P 25/08 Antiepileptics; Anticonvulsants
• CPC Class Codes: A61P 25/14 for treating abnormal movements, e.g. chorea, dyskinesia
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: A61P 25/28 for treating neurodegenerative disorders of the central nervous system, e.g. nootropic agents, cognition enhancers, drugs for treating Alzheimer's disease or other forms of dementia
• CPC Class Codes: A61P 43/00 Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
• CPC Class Codes: A61P 9/00 Drugs for disorders of the cardiovascular system
• CPC Class Codes: A61P 9/08 Vasodilators for multiple indications
• CPC Class Codes: A61P 9/10 for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis
• CPC Class Codes: C12N 2501/01 Modulators of cAMP or cGMP, e.g. non-hydrolysable analogs, phosphodiesterase inhibitors, cholera toxin
• CPC Class Codes: C12N 2501/105 Insulin-like growth factors [IGF]
• CPC Class Codes: C12N 2501/11 Epidermal growth factor [EGF]
• CPC Class Codes: C12N 2501/115 Basic fibroblast growth factor (bFGF, FGF-2)
• CPC Class Codes: C12N 2501/119 Other fibroblast growth factors, e.g. FGF-4, FGF-8, FGF-10
• CPC Class Codes: C12N 2501/13 Nerve growth factor [NGF]; Brain-derived neurotrophic factor [BDNF]; Cilliary neurotrophic factor [CNTF]; Glial-derived neurotrophic factor [GDNF]; Neurotrophins [NT]; Neuregulins
• CPC Class Codes: C12N 2501/135 Platelet-derived growth factor [PDGF]
• CPC Class Codes: C12N 2503/02 Drug screening
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 2510/00 Genetically modified cells
• CPC Class Codes: C12N 5/0618 : Cells of the nervous system
• CPC Class Codes: C12N 5/0623 : Stem cells
• CPC Class Codes: C12Q 1/6881 : for tissue or cell typing, e.g. human leukocyte antigen [HLA] probes
• CPC Class Codes: C12Q 2600/158 : Expression markers

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.

Claims

1. A method of screening a compound for an effect on a neural cell, comprising a) contacting a cell culture with a compound, wherein the cell culture comprises cells chosen from neuronal cells expressing neural cell adhesion molecule (NCAM) and glial cells, wherein the neuronal cells expressing NCAM or glial cells are progeny of a cell culture comprising cells expressing SSEA3, SSEA4, Tra-1-60 and Tra-1-81;

and b) determining any phenotypic or metabolic changes in the neuronal cells expressing NCAM or glial cells that result from contact with the compound thereby determining the effect of a compound on a neural cell.

2. The method of claim 1, wherein the cells are neuronal cells.

3. The method of claim 1, wherein the cells are glial cells.

4. A method of screening a compound for an effect on a neural cell comprising a) contacting a cell culture with a compound, wherein the cell culture comprises cells chosen from neurons expressing NCAM, astrocytes and oligodendrocytes, wherein the neurons expressing NCAM, astrocytes or oligodendrocytes, are progeny of a cell culture comprising cells expressing SSEA3, SSEA4, Tra-1-60 and Tra-1-81;

and b) determining any phenotypic or metabolic changes in the neurons, astrocytes or oligodendrocytes, that result from contact with the compound thereby determining the effect of a compound on a neural cell.

5. The method of claim 4, wherein the cells are neurons.

6. The method of claim 4, wherein the cells are astrocytes.

7. The method of claim 4, wherein the cells are oligodendrocytes.

8. A method for determining if a polynucleotide comprising a nucleotide sequence contained in an mRNA is more highly expressed in neural progenitor cells compared to cells expressing a marker chosen from GFAP and GalC, the method comprising:

a) determining the level of expression of a plurality of mRNAs in a neural progenitor cell population comprising cells expressing a marker chosen from NCAM and A2B5, wherein the cell population comprises cells that are the progeny of a cell culture comprising cells expressing SSEA3, SSEA4, Tra-1-60 and Tra-1-81;

b) determining the level of expression of a plurality of mRNAs in a cell population comprising cells expressing a marker chosen from GFAP and GalC; and

c) identifying an mRNA expressed at a higher level in the cells from a) compared to the ceils from b),

thereby determining if a polynucleotide comprising a nucleotide sequence contained in an mRNA is more highly expressed in neural progenitor cells compared to cells expressing a marker chosen from GFAP and GalC.