The aristaless related homeobox (ARX) transcription factor plays a crucial role in glucagon-producing alpha-cell differentiation. Here, we generate an ARX reporter iPSC line by 3' fusion of an intervening viral T2A sequence followed by a nuclear-localized histone 2B-cyan fluorescent protein (nCFP). The resulting cells have a normal karyotype and preserved pluripotency. In vitro differentiation of the ARX(nCFP/nCFP) reporter iPSCs towards the endocrine lineage confirmed the specific co-expression of the reporter protein in human glucagon' alpha-like cells. Thus, ARX(nCFP/nCFP) iPSC line will provide a powerful tool to monitor human a-cell progenitor differentiation as well as ARX(+) alpha-like cell function in vitro.