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Process for producing nerve stem cells, motor neurons, and GABAergic neurons from embryonic stem cells meetings achieved

Foreign code F110005211
File No. A192-02US
Posted date Aug 26, 2011
Country United States of America
Application number 47249001
Gazette No. 20040092012
Gazette No. 7294510
Date of filing Oct 3, 2001
Gazette Date May 13, 2004
Gazette Date Nov 13, 2007
International application number JP2001008703
International publication number WO2002081663
Date of international filing Oct 3, 2001
Date of international publication Oct 17, 2002
Priority data
  • P2001-099074 (Mar 30, 2001) JP
  • 2001WO-JP08703 (Oct 3, 2001) WO
Title Process for producing nerve stem cells, motor neurons, and GABAergic neurons from embryonic stem cells meetings achieved
Abstract (US7294510)
The present invention provides a method for producing motor neurons and GABAergic neurons characterized by including suspension-culturing embryonic stem cells in the presence or absence of a protein noggin to form embryoid bodies, selectively amplifying into neural stem cells from them by suspension culture in the presence of a fibroblast growth factor and a sonic hedgehog protein, and then differentiating the same.
According to this method, at least motor neurons and GABAergic neurons can be systemically and efficiently produced from ES cells.
Selective acquisition of neurons would be applicable to transplant therapy for amyotrophic lateral sclerosis, Huntington's chorea, Alzheimer's disease, etc.
Scope of claims [claim1]
1. A method for forming embryoid bodies, which comprises subjecting embryonic stem cells to suspension culture in the presence of a noggin protein, wherein said noggin protein is added to the culture prior to formation of embryoid bodies.
[claim2]
2. A method for producing neural stem cells, which comprises subjecting embryonic stem cells to suspension culture in the presence of a noggin protein, to thereby form embryoid bodies, and subsequently subjecting the embryoid bodies to suspension culture in the presence of a fibroblast growth factor and a sonic hedgehog protein to thereby form a culture of neural stem cells.
[claim3]
3. The method according to claim 2, wherein concentration of the fibroblast growth factor in culture medium is 5 to 50 ng/mL, and that of the sonic hedgehog protein in culture medium is 1 to 20 nM.
[claim4]
4. A method for producing motor neurons and GABAergic neurons, which comprises subjecting ES cells to suspension culture in the presence of a noggin protein, to thereby form embryoid bodies, and subsequently subjecting the embryoid bodies to suspension culture in the presence of a fibroblast growth factor and a sonic hedgehog protein, to thereby induce formation of neural stem cells, and differentiating the resultant neural stem cells in a differentiation medium.
[claim5]
5. The method according to claim 4, wherein concentration of the fibroblast growth factor in culture medium is 5 to 50 ng/mL, and that of the sonic hedgehog protein in culture medium is 1 to 20 nM.
[claim6]
6. The method according to claim 4, wherein the resultant neurons are substantially formed of motor neurons and GABAergic neurons.
[claim7]
7. The method according to claim 5, wherein the resultant neurons are substantially formed of motor neurons and GABAergic neurons.
[claim8]
8. The method according to claim 2, wherein the culture of neural stem cells comprises neurospheres.
  • Inventor, and Inventor/Applicant
  • OKANO HIDEYUKI
  • SHIMAZAKI TAKUYA
  • JAPAN SCIENCE AND TECHNOLOGY AGENCY
IPC(International Patent Classification)
Reference ( R and D project ) CREST Development, Differentiation, and Regeneration in Biological Systems AREA
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