Isolation And Use Of Pluripotent Stem Cell Population From Adult Neural Crest-Derived Tissues

Abstract

The present invention relates to methods of isolating a substantially homogenous population of pluripotent stem cells from adult neural crest tissue (e.g., periodontal ligament) as well as pharmaceutical compositions comprising such isolated pluripotent stem cells. Methods of inducing the isolated pluripotent stem cells into specific cell lineages, such as neurogenic and retinogenic lineages, are also described. The isolated pluripotent stem cells find use in various regenerative medicine applications and the treatment of degenerative diseases.

What is claimed is: 
     
         1 .- 26 . (canceled) 
     
     
         27 . A method of differentiating pluripotent stem cells isolated from adult periodontal ligament to neural progenitor cells or neural cells comprising:
 incubating said pluripotent stem cells in a culture media comprising epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) for a period of time, wherein a plurality of said pluripotent stem cells are differentiated into neural progenitor cells or neural cells at the end of the period of time.   
     
     
         28 . The method of  claim 27 , wherein EGF is present in the media at a concentration of about 25 ng/ml to about 150 ng/ml. 
     
     
         29 . The method of  claim 28 , wherein EGF is present in the media at a concentration of about 50 ng/ml. 
     
     
         30 . The method of  claim 27 , wherein bFGF is present in the media at a concentration of about 25 ng/ml to about 150 ng/ml. 
     
     
         31 . The method of  claim 30 , wherein bFGF is present in the media at a concentration of about 50 ng/ml. 
     
     
         32 . The method of  claim 27 , wherein the period of time is about 4 to about 12 days. 
     
     
         33 . The method of  claim 32 , wherein the period of time is about 8 days. 
     
     
         34 . The method of  claim 27 , wherein a plurality of the neural progenitor cells or neural cells express one or more markers selected from nestin, tubulin (TUBB3), neurofilament medium (NEFM), SOX1, synaptophysin, and glial fibrillary acidic protein (GFAP). 
     
     
         35 . The method of  claim 27 , wherein a plurality of the neural progenitor cells or neural cells exhibit a voltage-gated sodium current and/or a delayed, rectifying potassium current. 
     
     
         36 . The method of  claim 27 , wherein the neural cells are neurons and/or glia. 
     
     
         37 . The method of  claim 27 , wherein the pluripotent stem cells are human pluripotent stem cells. 
     
     
         38 .- 56 . (canceled)