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Pancreatic differentiation from human pluripotent stem cells

Item Type:Book Section
Title:Pancreatic differentiation from human pluripotent stem cells
Creators Name:Vinckier, N., Wang, J. and Sander, M.
Abstract:The ability to produce human pancreatic cells in vitro would open new possibilities for developing improved therapies through cell transplantation, disease modeling, and drug screening. Of particular medical importance are the insulin-producing beta cells of the pancreas, which are lost or dysfunctional in diabetes. Furthermore, an in vitro model of human exocrine cells could help devise new therapies for pancreatic exocrine disease, most notably pancreatic cancer. In the past decade much progress has been made in developing protocols to generate multipotent pancreatic progenitor cells from human pluripotent stem cells (hPSCs) that are capable of differentiating into both endocrine and exocrine cells. The sole approach that has proven successful is to reproduce essential steps of in vivo development in vitro through directed step-wise differentiation of hPSCs. The directed differentiation entails sequential exposure of hPSCs to different signaling factors, thereby moving cells through several developmental intermediates towards the pancreatic fate. Upon implantation into mice, hPSC-derived pancreatic progenitor cells spontaneously differentiate into endocrine and exocrine cells. Here, we describe a detailed protocol for the generation of pancreatic progenitor cells from hPSCs. We provide methods for the directed differentiation as well as the characterization of pancreatic progenitor cells and lineage intermediates by immunofluorescence staining and flow cytometry. With recently developed protocols, these pancreatic progenitor cells can be further differentiated in vitro into beta-like cells that functionally resemble immature human beta cells.
Keywords:Human Pluripotent Stem Cell (hPSC), Human Embryonic Stem Cell (hESC), Definitive Endoderm (DE), Gut Tube (GT), Posterior Foregut (FG), Pancreatic Endoderm (PE), SOX17, HNF4A, PDX1, SOX9
Title of Book:Working with stem cells
ISBN:978-3-319-30580-6
Publisher:Springer
Volume:1
Page Range:257-275
Number of Pages:374
Date:5 August 2016
Official Publication:https://doi.org/10.1007/978-3-319-30582-0_15

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