Clonal Derivation of Neural Stem Cells from Human Embryonic Stem Cells

Clonal culture is crucial for experimental protocols that require growth or selection of pure populations of cells. Currently, there is no method for deriving neural stem cells (NSCs) clonally from single human embryonic stem cells (hESCs). Bulk derivation of neural progenitors from hESCs for cell therapies can lead to a host of problems including incomplete differentiation leading to proliferation of tumorigenic clusters in vivo. Clonal derivation allows for the screening and selection of only the most suitable cells for culture and expansion. We have developed a clonal, serum free method of generating NSCs and their progenitors directly from hESCs with an efficiency of 1.6%. The NSC colony-forming cell was identified as a TRA-1-60/SSEA4 cell whose fate becomes specified in maintenance conditions by inhibition of bone morphogenic protein (BMP) signalling. This clonal culture method can be scaled up to produce vast quantities of NSCs for differentiation and use in cell therapies.