Abstract
Stepwise pancreatic β-cell differentiation protocols, designed to recapitulate key developmental milestones of pancreatic organogenesis-from definitive endoderm to mature, glucose-responsive β-cells-draw on insights from both rodent and human developmental biology. These protocols consist of three critical stages: the formation of definitive endoderm, the induction of pancreatic progenitors, and the maturation of functional β-cells. Here, we discuss human and rodent embryonic development together with stem-cell differentiation protocols in concert to identify gaps and bottlenecks that limit the practicality and scalability of current protocols. Ultimately, the aim of refining these processes is to produce functional β-cells from pluripotent stem cells to treat, and potentially cure, diabetes.