18-08-2016 дата публикации
Номер: US20160237404A1
Принадлежит:
The present invention discloses a method of identifying agents that affect human astrocytes functionality using ex-vivo differentiated pluripotent stem cells (PSC). In addition, the use of human progenitor astrocytes or human astrocytes for the treatment of Amyotrophic Lateral Sclerosis (ALS) in a human subject is also disclosed. 1. A method of screening an agent for preventing or treating Amyotrophic Lateral Sclerosis (ALS) the method comprising:(a) contacting a population of astrocytes, the astrocytes having been ex-vivo differentiated from pluripotent stem cells (PSC), with the agent;(b) co-culturing the population of astrocytes of step (a) or a conditioned medium thereof with a population of neurons; and(c) quantifying an effect of said agent to enhance survival or neural function of the population of neurons.2. The method of claim 1 , wherein said population of neurons is hypoxic claim 1 , under oxidative stress claim 1 , under glutamate toxicity or under AMPA/kainate toxicity.3. The method of claim 1 , wherein in step (b) the ratio of the population of astrocytes to neurons is greater than 1:1 claim 1 , 10:1 claim 1 , 100:1 claim 1 , 1000:1 claim 1 , or 10 claim 1 ,000:1.4. The method of claim 1 , wherein said astrocytes express each of GFAP claim 1 , GLAST claim 1 , AQP4 claim 1 , or a combination thereof.5. The method of claim 1 , wherein said astrocytes display secretion of neurotrophic factors selected from the group consisting of BDNF claim 1 , GDNF and VEGF.6. The method of claim 1 , wherein the oxidative stress is selected from the group consisting of reactive oxygen species (ROS) claim 1 , HO claim 1 , and any derivative thereof.7. The method of claim 1 , wherein said quantifying is conducted by counting the number of neurons which are under apoptosis.8. The method of claim 7 , wherein said apoptosis is detected by Caspase -3a labeling claim 7 , Annexin V claim 7 , Tubulin-B3 claim 7 , HB9 or DAPI.9. The method of claim 1 , wherein said agent is a ...
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