Cell Biology and Functional Genomics 1 article published in JoVE Biology Optimized Protocol for Generating Functional Pancreatic Insulin-secreting Cells from Human Pluripotent Stem Cells Ines Cherkaoui1, Qian Du2, Dieter Egli2, Shivani Misra1,3, Guy A. Rutter1,4,5 1Department of Metabolism, Digestion and Reproduction, Faculty of Medicine, Cell Biology and Functional Genomics, 2Departments of Pediatrics, Naomi Berrie Diabetes Center, Obstetrics and Gynecology, Columbia Stem Cell Initiative, Columbia University Irving Medical Center, Columbia University, 3Department of Diabetes, Imperial College Healthcare NHS Trust, 4Faculté de Médicine, Université de Montréal, 5Lee Kong Chian Imperial Medical School, Nanyang Technological University This article presents a protocol for directed differentiation and functional analysis of β-cell like cells. We describe optimal culture conditions and passages for human pluripotent stem cells before generating insulin-producing pancreatic cells. The six-stage differentiation progresses from definitive endoderm formation to functional β-cell like cells secreting insulin in response to glucose.
