Abstract
By virtue of their capability to replicate and regenerate, human stem-cell-derived beta-like cells could be a valuable resource for cellular therapy targeting insulin-dependent diabetes. Here, we present a protocol to generate beta-like cells from human embryonic stem cells (hESCs). We first describe steps for differentiation of beta-like cells from hESCs and CD9-negative beta-like cell enrichment through fluorescence-activated cell sorting. We then detail immunofluorescence, flow cytometry, and glucose-stimulated insulin secretion assay for characterization of human beta-like cells. For complete details on the use and execution of this protocol, please refer to Li et al. (2020).1.