Microglia derived from pluripotent stem cells and methods of making and using the same

Abstract

The present invention provides methods and compositions for the generation of microglial progenitor cells and microglial cells from pluripotent stem cells, such as embryonic stem cells and induced pluripotent stem cells. The present invention also provides cells produced using such methods, and both methods of treatment and methods of drug screening that use such cells. Also provided are various tissue culture media, tissue culture media supplements, and kits useful for the generation of human microglial progenitor cells and human microglial cells.

1 . A method for generating microglial cells comprising:
 a) culturing human pluripotent stem cells under conditions that induce myeloid differentiation, leading to the generation of CD14+ and/or CX3CR1+ microglial progenitor cells, and   b) culturing the CD14+ and/or CX3CR1+ microglial progenitor cells in either (i) a first microglial differentiation medium comprising IL-34, or (ii) a second microglial differentiation medium comprising M-CSF, thereby generating human microglial cells.   
     
     
         2 - 20 . (canceled) 
     
     
         21 . A method of treatment or prevention comprising administering microglial cells generated using the method of  claim 1  to a subject having, suspected of having, or at risk of developing a disease or disorder associated with a defect in or deficiency of microglial cells or microglial progenitor cells. 
     
     
         22 . The method of  claim 21 , wherein the disease or disorder associated with a defect in or deficiency of microglial cells or microglial progenitor cells is selected from the group consisting of amyotrophic lateral sclerosis (ALS), Alzheimer's disease (AD), multiple sclerosis (MS), Parkinson's disease, Rett syndrome, diffuse leukoenchephalopathy with spheroids, hereditary diffuse leukoenchephalopathy with axonal spheroids, frontotemporal lobar degeneration (FTLD), familial FTLD, schizophrenia, and autism spectrum disorders. 
     
     
         23 . The method of  claim 27 , wherein the microglial cells are generated from induced pluripotent stem (iPS) cells derived from somatic cells obtained from the subject. 
     
     
         24 . A method of identifying a compound useful in the treatment or prevention of a disease or disorder associated with a defect in or deficiency of microglial cells, the method comprising: contacting a microglial cell generated by the method of  claim 1  with a candidate compound, and determining whether the candidate compound improves the defect in or deficiency of microglial cells. 
     
     
         25 . The method of  claim 24 , wherein the method is a high-throughput method. 
     
     
         26 . The method of  claim 21 , wherein the subject is human. 
     
     
         27 . The method of  claim 21 , wherein the microglial cells are autologous. 
     
     
         28 . The method of  claim 21 , wherein the microglial cells are allogeneic. 
     
     
         29 . The method of  claim 28 , wherein the microglial cells are derived from a donor individual having an MHC/HLA type that matches that of the subject. 
     
     
         30 . The method of  claim 22 , wherein the disease or disorder associated with a defect in, or deficiency of microglial cells or microglial progenitor cells is Parkinson's disease. 
     
     
         31 . The method of  claim 22 , wherein the subject has a GBA N370S mutation.