Protocol to generate human pluripotent stem cell-derived pancreatic β cells through methionine and zinc deprivation

Efforts have been made to establish a differentiation protocol mimicking pancreatic development and to derive pancreatic β cells for regenerative medicine. Here, we present an optimized pancreatic β cell differentiation procedure using human pluripotent stem cells. We describe steps for a short 5-h methionine deprivation pretreatment followed by the application of zinc-deprived media at definitive endoderm differentiation stages to improve differentiation efficiency. The application of methionine and zinc deprivation facilitates the generation of functional pancreatic β cells. For complete details on the use and execution of this protocol, please refer to Sim et al. (2022).¹.