Methods of generating glial and neuronal cells and use of same for the treatment of medical conditions of the CNS

Patent metadata

Field	Value
Publication number	US-10258651-B2
Application number	US-201715468020-A
Country	US
Kind code	B2
Filing date	Mar 23, 2017
Priority date	Aug 28, 2006
Publication date	Apr 16, 2019
Grant date	Apr 16, 2019

Abstract

A method of generating neural and glial cells is provided. The method comprising growing human stem cells under conditions which induce differentiation of said human stem cells into the neural and glial cells, said conditions comprising the presence of retinoic acid and an agent capable of down-regulating Bone Morphogenic Protein activity.

First claim

What is claimed is:

1. A method of treating one or more symptoms of amyotrophic lateral sclerosis (ALS) in a subject in need thereof, the method comprising:
(a) culturing in suspension human embryonic stem cells in the presence of retinoic acid and Epidermal Growth Factor (EGF) to generate neurospheres;
(b) contacting the neurospheres with EGF on an adherent substrate, wherein the adherent substrate comprises an extracellular matrix component;
(c) dissociating the neurospheres following step (b) and contacting the dissociated neurospheres with a medium comprising EGF and Basic Fibroblast Growth Factor (bFGF);
(d) contacting the neurospheres following step (c) with a medium devoid of EGF and bFGF for 6-10 days, thereby generating astrocytes; and,
(e) directly transplanting the astrocytes into the central nervous system (CNS) of the subject.

2. The method of claim 1 , further comprising forming stem cell aggregates prior to step (a).

3. The method of claim 1 , wherein the concentration of EGF is 10-40 ng/ml.

4. The method of claim 1 , wherein the concentration of the retinoic acid is 1-50 μM.

5. The method of claim 1 , wherein step (a) is effected for 20-30 days.

6. The method of claim 1 , wherein the medium of step (c) comprises laminin and vitamin C.

7. The method of claim 1 , further comprising isolating the astrocytes following step (d).

8. The method of claim 1 , wherein the direct transplantation comprises an intrathecal injection of the astrocytes.

9. The method of claim 1 , wherein the subject is a human.

Assignees

• Yeda Res & Dev

Inventors

• Revel Michel
• Chebath Judith
• Izrael Michal
• Kaufman Rosalia

Classifications

• C12N2501/385

  of the family of the retinoic acid recptor, e.g. RAR, RXR; Peroxisome proliferator-activated receptor [PPAR] · CPC title

• C12N5/0619 Primary

  Neurons · CPC title

• C12N2506/02

  from embryonic cells · CPC title

• G01N33/5058

  Neurological cells · CPC title

• C12N2501/41

  Hedgehog proteins; Cyclopamine (inhibitor) · CPC title