In Vitro Generation of MGE Progenitors from Human Pluripotent Stem Cells
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Basal forebrain cholinergic neurons (BFCN) are required for memory and learning, and are affected in Alzheimer's disease (AD) causing cognitive deficits. The ability to selectively generate a source of BFCN from induced pluripotent stem cells (iPSC) would be a significant step for in vitro disease modeling and development of novel AD therapies. Efforts to produce BFCN include direct differentiation with overexpression of human transcription factors (LHX8 and GBX1) and using diffusible ligand (BMP9) (Bissonnette, 2011) and non-directed non-adherent differentiation protocol (NAdD) (Crompton, 2013). This current study focuses on defining whether the NAdD is superior to an adherent protocol in anterio-ventralization development of medial ganglionic eminence (MGE) cells, the progenitors of BFCN, and whether WNT inhibition would improve the efficiency of the NAdD protocol. Current findings provided evidence that NAdD derived cells expressed significantly higher levels of basal forebrain transcription factor (NKX2.1) compared to AdD define derived cells. By flow cytometry, NAdD has 20.8% and 9.9% more cells expressing PAX6 and FORSE1, respectively than AdD. The NAdD supplemented with WNT inhibition resulted in a significantly higher expression of NKX2.1 and yielded 39% more of FORSE1 positive cells than the classic NAdD. In conclusion, NAdD protocol is more efficient than the AdD protocol and WNT inhibition enhances the yield for MGE progenitors.