Figure 9:

Schematic overview of the experimental processes trialled to optimise the generation of vmDA neurons from human iNPs. Transfection of adult human fibroblasts with SOX2 and PAX6 followed by culture in reprogramming medium generates iNP cells that express SOX1, NESTIN, FOXG1, ASCL1 and NURR1, and can be differentiated into dopamine-like neurons positive for TH, AADC, VMAT2, GIRK2 and DAT. In order to enhance the yield and induce an authentic vmDA identity of these cells, a range of neural and dopaminergic lineage transgenes were trialled coupled with exposure to the patterning factors SHH-C24II, purmorphamine, CHIR99021 and/or FGF8 during reprogramming. The effect of exposure to the patterning factors SHH-C24II and FGF8 during the early stage of differentiation to promote a vmDA fate was also investigated.

AA = ascorbic acid.