Objective: Differentiation of hESCs to Dopaminergic neurons can be achieved in vitro by combination of small molecules during emerging neural ectoderm. Dopaminergic progenitors derived from hESCs are promising tool for the regenerative medicine in Parkinson cell therapy. Materials and Methods: To enable enrichment and characterization of Mesodiancephalon Dopaminergic (mDA) neural progenitors LMX1A locus of hESC (Royan H6 and Royan H5 cell lines from Royan Institute) targeted with GFP in the upstream of LMX1a coding sequence. GFP protein expressed as fused protein with LMX1a protein and both LMX1aGFP/GFP and LMX1aGFP/w clones were selected for differentiation and fluorescence-activated cell sorting (FACS) to select and purify midbrain dopamine progenitor cells. Initially, the dopaminergic marker profile of NPs cultures was evaluated after differentiation in vitro. Results: The overlap expression of eGFP with Lmx1a, LMX1b and Corin demonstrated that these cells were of a midbrain dopamine progenitor characters. In addition, enriched dopamine progenitors, which positively selected by FACS were viable, extended neurites, and maintained a dopaminergic profile in vitro, could give raise to mature DA neurons. Engraftment of GFP positive mDA progenitors and NCAM positive NPs as a control to rat model of Parkinson and behavior tests are under studies. Conclusion: We show that LMX1a could be used as a selection marker of early mDA progenitor’s in vitro and purified mDA will provide invaluable Materials for developmental studies and regenerative medicine.