Acceleration of stem cell differentiation

Abstract

Provide herein are compositions, methods and kits to accelerate pluripotent stem cell differentiation.

What is claimed is: 
     
         1 . A method to induce differentiation in stem cells, comprising,
 a) providing:   i) a cell culture comprising pluripotent stem cells,   ii) an inhibitor of Activin receptor-Like Kinase (ALK) 2/3 signaling,   iii) an inhibitor of fibroblast growth factor (FGF) signaling, or   iv) in place of or in combination with an inhibitor of FGF signaling, an inhibitor of Extracellular-Regulated Kinase (ERK) signaling,   v) an activator of WNT signaling,   b) exposing said pluripotent stem cells of i) simultaneously to culture medium containing inhibitors of ALK 2/3 of ii) and inhibitors of FGF of iii), and/or inhibitors of ERK signaling of iv); and activators of WNT signaling of v);   c) inducing differentiation of said pluripotent stem cells of b) into a population of cultured rostral hindbrain neuromesoderm expressing SOX2/Brachyury/GBX2/HOXB1.   
     
     
         2 . The method of  claim 1 , wherein said population of rostral hindbrain neuromesoderm further differentiates into cells selected from the group consisting of central and peripheral nervous system progenitor cells, patternable progenitor cells, neurons, glia and/or derivatives of the epidermis and neural crest, and somite. 
     
     
         3 . The method of  claim 1 , further comprising isolating said rostral hindbrain neuromesoderm. 
     
     
         4 . The method of  claim 1  further comprising converting said rostral hindbrain neuromesoderm into hindbrain neuroepithelium expressing SOX1/GBX2/HOXB1 comprising contacting said rostral hindbrain neuromesoderm with an inhibitor of fibroblast growth factor (FGF) signaling. 
     
     
         5 . The method of  claim 1  further comprising converting said rostral hindbrain neuromesoderm into cranial neural crest stem cells expressing PAX3/SOX10/SOX9/HOXB1 comprising contacting said rostral hindbrain neuromesoderm with an activator of bone morphogenic protein pathway. 
     
     
         6 . The method of  claim 4  further comprising converting said hindbrain neuroepithelium expressing SOX1/GBX2/HOXB1 into cranial neural crest stem cells expressing PAX3/SOX10/SOX9/HOXB1 comprising contacting said hindbrain neuroepithelium expressing SOX1/GBX2/HOXB 1 with an activator of bone morphogenic protein pathway. 
     
     
         7 . The method of clam 1, further comprising converting said rostral hindbrain neuromesoderm into caudalized neuromesoderm comprising contacting said rostral hindbrain neuromesoderm with an activator of WNT signaling, wherein the activator of WNT signaling contacts said rostral hindbrain neuromesoderm either alone or in combination with an activator of fibroblast growth factor receptor family. 
     
     
         8 . The method of  claim 7 , wherein said rostral hindbrain neuromesoderm can be converted into caudal hindbrain neuromesoderm expressing SOX2/Brachyury/HOXB4 within 24 hours with continued exposure to WNT/FGF signaling. 
     
     
         9 . The method of  claim 8 , wherein caudal hindbrain neuromesoderm can be converted into cervical spinal neuromesoderm expressing SOX2/T/HOXB4/HOXC5 within 24 hours with continued exposure to WNT/FGF signaling. 
     
     
         10 . The method of  claim 9 , wherein cervical spinal neuromesoderm can be converted into somite tissue by sustained WNT pathway activation in the absence of added FGF signaling. 
     
     
         11 . The method of  claim 8 , wherein said caudal hindbrain neuromesoderm is converted into cranial neural crest stem cells expressing PAX3/SOX10/SOX9 comprising contacting said caudal hindbrain neuromesoderm with an activator of SMAD signaling. 
     
     
         12 . The method of  claim 9 , wherein said cervical neuromesoderm is convened into trunk neural crest stem cells expressing PAX3/SOX10/SOX9 comprising contacting said cervical neuromesoderm with an activator of SMAD signaling.