Compositions and Methods for Enrichment of Neural Stem Cells Using Ceramide Analogs

• US 20090035285A1
• Filed: 09/22/2008
• Published: 02/05/2009
• Est. Priority Date: 09/25/2002
• Status: Abandoned Application

• Current Assignee: Medical College of Georgia (University System of Georgia), Bresagen, Inc. (Vertex Pharmaceuticals Incorporated)
• Original Assignee: Medical College of Georgia (University System of Georgia), Bresagen, Inc. (Vertex Pharmaceuticals Incorporated)
• Inventors: Condie, Brian G., Bieberich, Erhard
• Application Number:
  US12/234,885
• Publication Number:
  US 20090035285A1
• Time in Patent Office: Days
• US Class Current: 424/93.700
• CPC Class Codes: A61P 25/16 Anti-Parkinson drugs
• CPC Class Codes: C12N 2500/34 Sugars
• CPC Class Codes: C12N 2500/36 Lipids
• CPC Class Codes: C12N 2500/90 Serum-free medium, which may still contain naturally-sourced components
• CPC Class Codes: C12N 2502/14 hepatocytes
• CPC Class Codes: C12N 2506/02 from embryonic cells
• CPC Class Codes: C12N 5/0618 Cells of the nervous system
• CPC Class Codes: C12N 5/0619 Neurons

Abstract

The present invention provides compositions and methods for human neural cell production. More particularly, the present invention provides cellular differentiation methods employing amphiphilic lipid compounds, preferably ceramide analogs of the β-hydroxyalkylamine type and optionally employing an essentially serum free MEDII conditioned medium for the generation of human neural cells from pluripotent human cells. The methods alternatively comprise modulating apoptosis by modifying the levels of PAR-4, with or without the presence of amphiphilic lipid compounds and optionally employing MEDII conditioned medium. The methods alternatively encompass modulating apoptosis by modulating the intracellular concentration of endogenous lipid second messengers, such as ceramide.

Claims

1-23. -23. (canceled)

24. :

A method of producing a human neural cell comprising,

a) providing a pluripotent human cell,

b) forming an embryoid body comprising the pluripotent human cell by culturing the pluripotent human cell in a medium that is optionally essentially serum free; and

c) culturing cells from the embryoid body with a composition comprising a sphingosine compound of the general formula

25. :

The method of claim 24, wherein the medium is an essentially serum free medium and comprises a MEDII conditioned medium.

29. A human neural cell produced by the method of claim 24.

30. :

A method for treating a patient, comprising a step of administering to the patient having a neural disease a therapeutically effective amount of the neural cell of claim 29.

31. :

The method of claim 30, wherein the neural disease is Parkinson'"'"'s disease.

26. :

A method of producing a human neural cell comprising,

a) providing a pluripotent human cell;

b) forming an embryoid body comprising the pluripotent human cell by culturing the pluripotent human cell in a medium that is optionally essentially serum free; and

c) culturing cells from the embryoid body with a composition comprising a hydroxyalkyl ester compound of the general formula

27. :

The method of claim 26, wherein the medium is an essentially serum free medium and comprises a MEDII conditioned medium.

28. :

The method of claim 27, wherein the composition comprises an hydroxyalkyl ester compound of the structure

72. :

A human neural cell produced by the method of claim 26.

73. :

A method for treating a patient, comprising a step of administering to the patient having a neural disease a therapeutically effective amount of the neural cell of claim 72.

74. :

The method of claim 73, wherein the neural disease is Parkinson'"'"'s disease.

32-71. -71. (canceled)