Establishment of Vietnamese human induced pluripotent stem cell-derived Orexin-A neurons

Abstract

Narcolepsy type 1 is a neurological disorder that severely affects sleep control due to a decrease in orexin neurons. Although research on neurological diseases, including narcolepsy, is vital for advancing science and improving human life, progress is hindered by the challenge of obtaining human brain samples as well as the difficulty in culturing neuronal cell lines. To overcome this obstacle, we generated Orexin-A neurons from human induced pluripotent stem cells (hiPSCs) by utilising a differentiation medium supplemented with N-acetyl-D-mannosamine (ManNAc) and Bone morphogenetic protein 4 (BMP4). The cells obtained from our study were characterised using immunofluorescence staining, real-time PCR, and competitive ELISA technique. The results show that our research successfully generated a hiPSC line with pluripotent stem cell characteristics. Furthermore, Orexin-A neurons derived from hiPSCs showed positive results for NCAM1 and TUBB3 markers, an increase in HCRT gene expression, and the capacity to secrete Orexin-A. These promising findings suggest that our hiPSC-derived Orexin-A neurons could serve as a foundation for cell-based disease modelling and further investigations into the underlying mechanisms of related neurological disorders.