Development of a motor neurons-specific fluorescence reporter system

Abstract:

Human induced pluripotent stem cells (hiPSCs) have the potential to differentiate into multiple cell types. Motor neurons (MNs) differentiated from hiPSCs are important models of many motor neuron diseases. To simplify the identification of MNs, lentivirus vectors were used to transfer MNs-specific promoter HB9 and red fluorescent protein (RFP) gene into hiPSCs-derived human neural stem cells (hNSCs). Stable positive cells hNSCs-HB9-RFP-Puro were obtained after antibiotic selection. Subsequently, the positive cell line was infected with lentiviruses LV-Ngn2-Sox11-GFP and LV-Isl1-Lhx3-Hygro, which overexpressed the MNs differentiation transcription factor, and differentiated to MNs directly. Differentiated mature MNs showed neuron-like structure, expressed RFP and neuron-related markers β-tubulin and choline acetyltransferase (ChAT) under the control of the MNs-specific promoter HB9. The fluorescence reporter system provides a visual method for directed differentiation and identification of MNs, and may promote the applications of MNs in disease models and drug screening.